life science - eurogentec · pdf filepage 5 genomic solutions life science oligonucleotides...

OligonucleotidesProviding you more…

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General information

Eurogentec S.A., part of Kaneka Corporation, is a leading global supplier of innovative reagents, kits, specialty products and custom services. Through its three inter-related business units (BU), the company provides high quality products to scientists involved in the life science, biotechnology, diagnostic and pharmaceutical markets.

Eurogentec is fully ISO 9001, ISO 13485 certified, cGMP accredited by the Belgian Ministry of Health and approved by the US FDA for the commercial manufacturing of a biologic for the US market.

EGT Group (including AnaSpec and EGT North America) currently employs around 325 people, including more than 40 PhDs.

Kaneka Corporation is an innovation-oriented chemical company. Business activities span a broad spectrum of markets ranging from plastics, EPS resins, chemicals and foodstuffs to pharmaceutical intermediates, medical devices, electrical and electronic materials and synthetic fibers.

Kaneka Corp. is a company headquartered in Osaka-Japan, with subsidiaries in Belgium, the United States, Singapore, Malaysia, China, Australia, Vietnam, India, South Korea and Taiwan.

Kaneka employs more than 8400 people worldwide. n

The In Vitro Diagnostics BU provides technical and project support for contract manufacturing of GMP oligonucleotides and Taq DNA polymerases for use in Molecular Diagnostic applications. We also produce pre-dispensed custom assays in a GMP environment n

The GMP BioManufacturing BU is a full-service Contract Manufacturing Organization (CMO) and offers significant know-how in components manufacturing. Eurogentec is a full service CMO specialized in the production of biologics. We also offer significant experience in process development and manufacturing of recombinant proteins and protein conjugates from E. coli, P. pastoris and other microbes, as well as plasmid DNA manufacturing. n

The Life Science BU is specialized in innovative solutions in Genomics and Proteomics. Eurogentec proposes a wide range of reagents, kits and consumables for Life Science applications including oligonucleotides, (q)PCR kits and reagents, high quality AnaSpec® Peptides, Custom and Catalogue antibodies and many more).n

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Table of contents 2 GeneralInformation 4 Features&Benefits 6 LifeScienceOligonucleotides 8 Oli&GO™10 CustomOligonucleotides14 Real-TimeqPCRProbes17 RNAiOligonucleotides19 UniversalPrimers20 PNAFishProbes21 UniqueOligonucleotides22 MinimumGuaranteedYields23 Shipping24 Documentation25 OrderingInformation26 TipsandTricks28 RelatedProducts/andMuchMore29 TrademarkLabels30 LicenseStatements

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Features & benefits

Custom Products¡ AlltypesofOligonucleotides,

from2to225bases¡ Allchemistries:DNA,RNA,LNA®,

2’O-Me,PNA¡ Morethan300modifications¡ Allsynthesisscales,fromµgtograms¡ WiderangeofReal-TimeqPCRProbes¡ RNAiOligonucleotidesTrusted quality¡ Optimizedchemistry¡ StringentQualityControls¡ ISO9001CertifiedQualitySystem¡ ISO13485CertifiedforIVDOligonucleotides

True partnership¡ Fastturnaroundtimes¡ Technicalandscientificsupport,staffedby

qualifiedandexperiencedscientists,inMolecularBiologyandChemistry

Reduced environmental footprint¡ OptimizedOligonucleotidessynthesis¡ PaperfreeOligonucleotiderelated

documentation¡ Dryicefreeshipmentsinrecycledpaper

packagings

Dependingonthefieldofapplications(Research&Development,Clinical

MolecularDiagnosticassay,MolecularDiagnosticcommercialkit),EurogentecprovidesdifferentgradesofOligonucleotideswithdifferentQA/QCandtraceabilityrequirementsappropriatetotheseapplications.n

4ManufacturingSites(LiègeBE,CamberlyUKandFremontUSA)SalesOfficesandDistributors

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Life Science Oligonucleotides

OurLifeScienceOligonucleotideserviceoffersalargevarietyofsynthesisscales,morethan300modificationsincludinghighlymodifiedOligonucleotides,sophisticatedchimeras,siRNAcoupledtovariouspeptides,OligonucleotidescontainingmultipleG-stretches,aptamers…

30yearsofexperience

Anextensiverangeofpurifications,smallandlargescale,formatsandpackagingsarealsooffered.TheLifeScienceOligonucleotidesareproducedunderISO9001qualitystandard.n Custom

Research OligonucleotidesLarge Scale OligonucleotidesTrack OligonucleotidesPre-Diagnostic OligonucleotidesDiagnostic Oligonucleotides

EUROGENTEC IS A 30-YEAR LEADING SUPPLIERofhigh-qualityreagentsandcustom-synthesizedOligonucleotidestoscientistsaroundtheglobe.

cGMP Oligonucleotides for Commercial kitscGMP Oligonucleotides ensure exceptional product quality by manufacturing in classified cleanrooms and use

of an ISO 13485-certified and GMP compliant QMS. This unique level of compliance ensures a high standard of quality, exceptional lot-to-lot reproducibility and fully documented traceability including comprehensive batch

records upon final QC release of products. Eurogentec is also registered with the FDA as a Class I manufacturer of custom ASR (Analyte Specific Reagent) Oligonucleotides, defined by the FDA as being intended for use in

in vitro diagnostic (IVD) applications.

Track OligonucleotidesTrack Oligonucleotides are non-IVD Oligonucleotides offering a higher traceability (and other quality assets) in

the production process than RUO Oligonucleotides. The traceability includes identification of the equipment used, the personnel involved and other important manufacturing specifications.

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Life Science OligonucleotidesWHATEVERyour

application,eventhemostdemandingones(NMR,

X-raycrystallography,in vivouse…),Eurogenteccanprovidethehighest

qualityOligonucleotidestomeet(andexceed)your

expectations!

*Largersynthesisscalesareavailableonrequest.

www.eurogentec.com/oligonucleotides.html

s PCR Primerss Double-Dye Probess LNA® Double-Dye Probess MGB Probess Molecular Beaconss LC Hybridization Probess Plexor™ Primerss NASBA®

s MLPA

Real-Time qPCR Probes

Length From 8 to 50 bases

Synthesis scale 10 nmol • 40 nmol • 200 nmol • 1000 nmol • 2.5 µmol

• 5 µmol • 10 µmol*

Backbone DNA, LNA®, 2’O-Me RNA and Phosphodiester linkages

Modifications 5’: 6-FAM, HEX, Cy®3, TET , Cy5® 3’: TAMRA, DABCYL,

BHQ®, DDQ…

Purifications RP-HPLC or Dual HPLC

Quality Control MALDI-TOF MS and Analytical HPLC

Format Dried

Packaging 2 mL tube

Documentation Technical Data Sheet

Probe Design Available on request

Shipping At Room Temperature

Custom Oligonucleotides


Synthesis scale 10 nmol • 40 nmol • 200 nmol

• 1000 nmol • 2.5 µmol • 5 µmol • 10 µmol*

Backbone DNA, RNA, LNA®, 2’O-Me RNA, PNA and

all linkages

Modifications More than 300 modifications!

Purifications SePOP Desalting, RP-Cartridge•Gold™, HPLC,

PAGE, Dual HPLC or UltraPureGold™

Quality Control MALDI-TOF MS

Format Dried (except for unmodified SePOP Desalted

Oligonucleotides from 15 to 39 DNA bases:

100 µM H2O)

Packaging 2 mL tube, 96-well or 384-well plates



s PCR Primerss FISH Probess Pyrrosequencing s Primerss Cloning Linkerss Antisense Oligonucleotidess Highly Modified Oligonucleotides

NGS Oligonucleotides


Delivery quantity 10 nmol minimum

Backbone DNA and Phosphorothioate linkage

Modifications 5’Phosphate and 5’Biotin-TEG

Purifications RP-HPLC or RP-Cartridge


Format Dried

Packaging 2 mL Tube


Shipping Free shipping at Room Temperature

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¡DuringOligonucleotidesynthesis,eachnucleotideis

coupledsequentially(from3’to5’)tothegrowingchainfollowinga

synthesiscyclebasedonstandardß-cyanoethylchemicalreaction:

deblocking,coupling,capping,oxidationanddeblocking.AttheendoftheOligonucleotidesynthesis,the

crudeproductiscleavedfromthesolidsupport(CPGorpolystyrene

beads).n

Oligo Centre 00 800 666 00 123European toll free number

[email protected] www.eurogentec.com

Universal Primers

Final Yield 1 OD/5 nmol

Backbone DNA

Modifications None

Purifications RP-HPLC

Quality Control MALDI-TOF MS and CGE

Format Dried

Packaging 2 mL tube



s SP6 Promoter s T7 terminator s M13s Bluescript SKs pET3's oligo dT 16 Bases

s Telomere Cs Telomere G

PNA FISH Probes

Length 18 bases

Synthesis scale 5 nmol

Backbone PNA

Modifications FAM, Cy®3, FITC, Cy5®, TMR

Purifications RP-HPLC

Quality Control MALDI-TOF MS + RP-HPLC

Format Dried

Packaging 2 mL tube


Shipping At room temperature

s Your Oligonucleotides

Unique Oligonucleotides


Synthesis scale Customized

Backbone Usual and atypical Chemistry

Modifications Common and rare modifications

Purifications SePOP Desalting,

RP-Cartridge•Gold™, HPLC, PAGE, Dual HPLC

or UltraPureGold™

Quality Control Adapted to your needs

Format Adapted to your needs

Packaging Adapted to your needs

Documentation Technical Data Sheet and Custom Documentation


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SynthesisCycle

1. Deblocking

2. Coupling

3. Capping

4. Oxidation

RNAi Oligonucleotides


Synthesis scale 10 nmol • 40 nmol • 200 nmol

• 1000 nmol*

Backbone RNA, LNA®, 2’O-Me RNA

and all linkages

Modifications 5’:Phosphate, 6-FAM, Cy®3, Cy5®, TET,

HEX,... 3’:DABCYL, TEG-Cholesteryl,

TAMRA...

Purifications SePOP Desalting, RP-HPLC or PAGE


Format Dried

Packaging 2 mL tube


siRNA Design Free and guaranteed


s Custom siRNA Duplexes s Control siRNA Duplexes s miRNA Inhibitors s miRNA Mimics

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Oli&GOTM

¡ Secure: –Usermanagement –Scheduledorders¡ Attractive: –Exclusivepricing¡ Convenient: –1invoiceformultipleorders –Real-timeaccountstatus –Comprehensiveorderhistory

www.eurogentec.com/oli-and-go.html

A unique e-commerce platform for all your oligos

Your budget under control

Manage your orders and your key users

Catalog Numbers

FunctionalitiesUse1ormoreAccounts ✔ ✔

BuyOligonucleotides ✔ ✔

ReceiveanOrderConfirmation ✔ ✔

ReceivetheRelatedDocumentation ✔ ✔

RenametheAccount ✔ ✘

Add/RemoveUser(s)* ✔ ✘

Define/UpdateShippingAddress ✔ ✘

ReloadtheAccount(s) ✔ ✘

ScheduleOrders(Day/Time)NEW ✔ ✘

*Multipleuserscanbedefinedperaccount

USERADMINISTRATOR Fullofprivilegestocontrol

thesystem.Restrictedaccess.

Description Cat.#Activation

Oli&GOTM Activation XS CD-OG002-XS

Oli&GOTM Activation M CD-OG002-M

ReloadOli&GOTM Reload S CD-OGR02-S

Oli&GOTM Reload M CD-OGR02-M

Oli&GOTM Reload L CD-OGR02-L

Oli&GOTM Reload XL CD-OGR02-XL

For pricing information please refer to the Eurogentec price listTo get a quotation, please contact us at:[email protected]

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Focusonyoursequenceonly.

GO back to bases GE

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A unique e-commerce platform for all your oligos

Your budget under control

Manage your orders and your key users

Account #1Project,Grant,Team,Lab,

Department,Institute…

Account #2Project,Grant,Team,Lab,

Department,Institute…

ADMINISTRATORBuyer,LabManager,

Scientist,PhD,Technician

User1 User1

User2 User2

User3 User4

UserN UserN

Scientist,PhDStudent,Technician

Exampleofkeyusermanagementwith1administratoraccountand2useraccounts.Eachuseraccountcanbeusedbymultipleusers.Adefineusercanusemultipleuseraccounts.

Scientist,PhDStudent,Technician

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BasesDNA Oligonucleotides are used in many applications such as PCR and DNA array…RNA Oligonucleotides are particularly suitable to study the involvement of RNA in cells regulation.LNA® Oligonucleotides possess high thermal stability and are recommended for hybridization assays requiring high specificity.2’O-Me RNA Oligonucleotides are very stable and nuclease resistant, thus recommended for antisense studies.PNA is artificial DNA/RNA analogue with no charged backbone. PNA is particularly useful for FISH studies, miRNA inhibition…

Length From2to139basesSynthesis scale 10nmol•40nmol•200nmol

•1000nmol•2.5µmol•5µmol•10µmol*

Backbone DNA,RNA,LNA®,2’O-MeRNA,PNAandalllinkages

Modifications Morethan300modifications!Purifications SePOPDesalting,

RP-Cartridge•Gold™,HPLC,PAGE,DualHPLC,UltraPureGold™

Quality Control MALDI-TOFMSFormat Dried(exceptforunmodifiedSePOP

DesaltedOligonucleotidesfrom15to39DNAbases:100µMH2O)

Packaging 2mLtube,96-wellor384-wellplatesDocumentation TechnicalDataSheetShipping AtRoomTemperature


*OnlyavailablewithDNAbasesLNA®

Backbones

LinkagesPhosphodiester (default) bonds connect the 3' carbon atom and the 5' carbon of the sugar.Phosphorothioatebonds possess an increased resistance against nuclease due to the substitution of a non-bridging oxygen by sulphur. Methylphosphonate* bonds are non ionic nuclease resistant linkages. n

SINCE 1987, Eurogentec’smissionhasbeentoansweryourneedsinOligonucleotideswhateveryourapplication.

From4elementarymodulesbuildandcustomizeyourOligonucleotideswithallyourdesiredspecifications.

k Backbones:Fivedistincttypesofbases(DNA,RNA,LNA®,2’O-MeandPNA)and3differentlinkages(Phosphodiester,MethylphosphonateandPhosphorothiate)areavailable.

k Modifications:Withacataloguerichofmorethan300modifications,youwillfindthemostcommonaswellasthemostunsualcombinationforyouroligonucleotides.e.g.:N4-ethylanalogue,2-Aminopurine,AP-dC,...

k Purifications:Dependingtoyourapplication,alargevarietyofoligonucleotidepurificationmethodsareproposed.

k Additional services:Oligonucleotideformat(dried,insolution,mixedandannealed)andpackaging(96or384wellplatesandaliquotingoption)maybecustomisedtoperfection.AdditionalQualityControl(MALDI-TOF,HPLC,CGE…)givingqualitativeand/orquantitativeinformationontheoligonucleotidepurificationcanbeperformedonrequest.n

We offer various chemistries as backbone (DNA, RNA, LNA®, 2’ O-Me and PNA) each bone can be linked to the next one by phosphodiester, methylphosphonate or phosphorothiate linkages.

O

O O

O O'

BaseO

LNA®

2'O-Me RNA

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R = A/G Y = C/T M = A/C K = G/T

S = C/G W = A/T B = C/G/T D = A/G/T

H = A/C/T V = A/C/G N = A/C/G/T

Phosphates5’ Phosphate 3’ Phosphate

QuenchersTAMRA, DABCYL, Eclipse® Dark Quencher, Deep Dark Quenchers,

BHQ®

Biotins5’ Biotin TEG, Internal Biotin,

3’ Biotin, dR Biotin

Modifiers Amino Modifiers C3, C6, C12,

Thiols Modifiers

FluorophoresFAM, Fluorescein, HEX, TET, JOE, ROX, Yakima Yellow®, Dragonfly

Orange™, Cy®3, Cy®5, Alexa Fluor®,…

Degenerate BasesWobbles & Spikes

Non-Natural Bases Inosine, Universal bases, IsodC-IsodG, C5-propyne and methyl

analogues,…

Spacers Spacers C3, C9, C12,…

Conjugates Digoxigenin, Dinitrophenol,

Cholesteryl, Psoralen, Carboxy,…

Modification(s)

OverviewOligonucleotides can be modified by direct incorporation during the synthesis or post-synthesis labelling.

Direct incorporation3’ modificationsSince automated oligonucleotide synthesis is realized from 3’ to 5’, these modifications are only possible if the corresponding solid support (CPG column) is available and if the modification is compatible with the chemistries used during the synthesis. Typical examples are 3’-phosphate, 3’ Biotin, 3’ FAM, 3’ DDQ I, 3’ BHQ-1®…

5’ and internal modificationsMany modifications can be directly introduced at the 5’ end or at internal positions of the Oligonucleotides using the phosphoramidites. However, these modifications need to support the somewhat harsh cleavage-deprotection conditions including a strong basic pH. Typical examples are 5’ Biotin, 5’ Phosphate, 5’ Cholesterol, 5’ FAM, 8-Oxo-dA, Biotin-dT, DABCYL-dT…

Post-synthesis incorporationThe two major post-synthesis reactions illustrated below are used to introduce sensitive dyes or compounds that do not exist as phosphoramidites. In the first case, the label is conjugated to an amino-modified oligonucleotide (3’, 5’ or on a dT) using its amino-reactive version (N-hydroxysuccinimide (NHS) ester in most cases).

Whatever your application, even the most specific ones (SNP, FISH, RNAi...), Eurogentec can provide the most exotic modifications according to your application needs !

A second possibility (originally also used for synthesis of molecular beacons) is the addition of a maleimide-modified label to a thiol-modified oligonucleotide. n

IUB base codes

More than 300 Modifications!

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Purification(s)

The initial aim of any purification method is to remove the by-products resulting from the removal of the protecting groups and other synthesis by-products. As with the purification of other biomolecules, the removal of small inorganic impurities is referred to as “desalting”.

In general, the purity level required for a specific application depends on the potential downstream problems, which may occur as a result of the presence of those truncated sequences. This purification step ensures that even the simplest primer will be suitable for most molecular biology applications, such as PCR, RT-PCR, Real-Time qPCR, sequencing and hybridization studies.

If you hesitate between purifications, than specify ”Recommended Single Purification" (additional fee) on your order and we will choose for you the best purification adapted to your oligonucleotide. n

Applications

NMR, X-ray crystallography

Production of cloning linkersSite-directed mutagenesis

Cloning and subcloning PCRGene synthesisGel-shift assay

Special modifications (G-clamp...)miRNA, siRNA and antisenseFirst-strand cDNA synthesis

in situ HybridizationReal-Time qPCRCapillary sequencingNon-radioactive labelling

AFLPOLASensitive PCR (Diagnostic)Classical modifications(modified bases, chemical linkers...)

Isothermal sequencingCycle sequencingRoutine PCRHybridizationDNA MicroArraySNP Analysis

>95 %

% o

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l len

gth

Olig

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ides

*

*These values are purely indicative and only valid for an unmodified

Oligonucleotide of 20 bases. In addition, according to your Oligonucleotides

(sequences, modifications…), the purity level can be analysed by various methods

(analytical HPLC, CGE…).

70 %

Length bases

UltraPureGold™

PAGE

Dual HPLC (RP+RP)

Dual HPLC (RP+IEX)

HPLC (RP or IEX)

SePOP desalting

RP Cartridge • Gold™

5-9 10-39 40-59 60-79 80-99 100-139

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Additional service(s)

MALDI-TOF Mass Spectrometry:Thismethodprovidesthemostpreciseinformationaboutthelength,deprotection-productandverificationofthepresenceoflabelsformodifiedOligonucleotidesoverabroadrangeoflengths(upto60bases).

RP-HPLC:ThisisaveryefficienttechniqueprovidingyouquantitativeinformationaboutthepuritylevelofOligonucleotidesfrom15to40baseslong.

IEX-HPLC:ThistechniqueisparticularlyadaptedtoquantifythepuritylevelofOligonucleotidesfrom15to40baseslong.

Capillary Gel Electrophoresis (CGE):ThismethodisadaptedtoassessverypreciselythepurityofOligonucleotideslongerthan40bases.

Fluorescence analysis: Thisnon-destructivephysicaltechniqueprovidesqualitativeinformationaboutyourfluorescentOligonucleotides.n

Dried: AllthesynthesisedOligonucleotidesaredriedbydefault(exceptSePOPunmodifiedOligonucleotidesfrom15to39bases).

In solution:Youmayselectthenatureofthereconstitutionbuffer(H2OorTE),thevolumeofthereconstitutionbuffer(from50to1000µl)or/andthefinalOligonucleotidesconcentration(from5to250µM).

Annealed: siRNAorcloninglinkersareannealedbydefault.

Mixed: SimilaramountsofforwardandreverseOligonucleotidescanbemixedinasingletube.n

2 mL tube: Bydefault,eacholigonucleotideisprovidedinindividual2mLtube.

96-well plates: Clustertubes,wellplateanddeepwellplateareavailable.

384-well plates: Speciallysuitableforhighthroughputexperimentsrequiringupto96Oligonucleotides.

Aliquoting: AlltheOligonucleotidesinsolutioncanbesplitinsmallaliquotsofdesiredvolume(from50to1000µl).n

YourOligonucleotidescanbeexpressshippedin24hoursuponrequestforordersupto24unmodifiedSePOPDesalted(100µMH2O)andRP-Cartridge•Gold™(Dried)CustomOligonucleotides(15-30bases/10-40nmolscale).n

Wecontinuouslyupdateoursoftwareanddesignrulestoreflectthelatestscientificdevelopmentsaswellasintegratecustomerrequirements.Thisserviceincludesprimers,Double-DyeOligonucleotides,MolecularBeacons,siRNAdesign,miRNAinhibitors...n

Additional QC Format Packaging Shipping Design

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Real-Time qPCRprobes


•1000nmol•2.5µmol•5µmol•10µmol*

Backbone DNA,LNA®,2’O-MeRNAandPhosphodiesterlinkage

Modifications 5’:6-FAM,HEX,Cy®3,TET,Cy5®…3’:TAMRA,DABCYL,BHQ™,DDQ…

Purifications RP-HPLCorDualHPLCQuality Control MALDI-TOFMSandAnalyticalHPLCFormat DriedPackaging 2mLtubeDocumentation TechnicalDataSheetProbe Design AvailableonrequestShipping Atroomtemperature


EUROGENTEC OFFERS a wide range offluorophores and quenchers in variouscombinations to fit any methods and Real-Time thermocyclers. If any help is neededconcerning the combination of dyes thatshould be used on a specific thermocycler,pleasedonothesitatetocontactourscientificsupportthatwillbehappytohelpyou.n

Double-Dye probesDouble-DyeProbeshaveafluorescentreporterdyeandaquencherattheir5’and3’ends,respectively.Duringtheamplificationprocess,the5’p3’exonucleaseactivityoftheTaqDNApolymerasecleavesthefluorophorefromtheprobe.SincethefluorophoreisnolongersubjectedtoFRETquenching,itstartstofluoresce.Thisfluorescencecanbemeasured,andthelevelisdirectlyproportionaltotheamountoftargetDNAaccumulatingduringthePCRreaction.

LNA® Double-Dye probesLNA®baseshaveamodificationtotheribosebackbonethatlocksthebaseintheC3’-endoposition,whichfavors

RNAA-typehelixduplexgeometry.ComparedtoDNADouble-Dyeprobes,LNA®Double-Dye

probeshaveseveraladvantages:k TheyexhibitunprecedentedthermalstabilitiestowardscomplementaryDNAandRNA.k LNA®basesshowbettermismatchdiscrimination.k Thehigh-bindingaffinityofLNA®Oligonucleotidesallowstheuseofshorterprobes.k Theyofferhighspecificityand/orreproducibility.k Furthermore,LNA®offersthepossibilitytoadjustTmvaluesofprimersandprobesinmultiplexassays.n

Increase the stability of

your Duplex

Double-Dye Probes

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MolecularBeaconsareprobeswhichcontainastem-loopstructure,afluorophoreandaquencherattheir5’and3’ends,respectively.Inthepresenceofacomplementarysequence,theprobeunfoldsandhybridizestothe

target,thefluorophoreisdisplacedfromthequencher,whichcannolongerabsorbthephotonsemittedbythefluorophore,andtheprobestartstofluoresce.Theamountofsignalisproportionaltotheamountoftargetsequence,andismeasuredinReal-Timetoallowquantificationoftheamountoftargetsequence.EurogentecisalicensedsupplierofMolecularBeacons,andoffersalargenumberoffluorescentreportersandquenchers.

Wavelength-Shifting Molecular BeaconsWavelength-ShiftingMolecularBeaconsarebrighterthanstandardMolecularBeaconsbecauseofanenhancementintheintensityofthefluorescenceoftheemitterfluorophore.

2’ O-Methyl RNA Molecular Beacons2’O-MethylRNAprobesareconsideredtoperformbetterthenDNAOligonucleotidesbecausetheyarenotonlynucleaseresistant,butalsopossessahigheraffinity,anincreasedspecificity,afasterhybridizationkinetics,andasuperiorabilitytobindtostructuredtargetscomparedtoDNAOligonucleotides.n

MGBincreasestheTmofaprobebecauseofitsminorgroovebindingability.MGBProbesformhighlystableduplexeswiththeirtargets

Enhance your

detection

Molecular Beacons

FRET probes to quantify

target sequences

High quality MGB probes

LChybridizationprobes,alsocalledFRETprobes,arelabelledwithasinglefluorescentreporter.TwoLChybridizationprobesspecificfortwoadjacentsequencesinthetargetDNAareusedforquantitativeReal-TimePCR.Oneprobeislabelledwithadonorfluorophoreandtheotherlabelledwith

anacceptor(orreporter)fluorophore.Whentheprobesareboundtothetargetsequence,thefluorescentsignalistransferredfromthedonortotheacceptor,whichstartstofluoresce.Whenprobesarenotannealedonthetargetsequence,donorandacceptordyesarenotincloseproximityandnoemissionwasdetected.Theamountofsignalisproportionaltotheamountoftargetsequence,andismeasuredinrealtimetoallowquantificationoftheamountoftargetsequence.A3’phosphategroupisalsoaddedtopreventextensionofthereporterprobebyTaqDNApolymeraseduringthePCRcycles.n

allowingshorterprobestobehighlyefficient.ThereforeMGBprobesaremorespecific,moreefficientandmoresensitivethanstandarddouble-dyeprobes.Relyingona30-yearexpertiseinoligonucleotidemanufacturing,EurogentecnowprovideshighqualityMGBprobesperfectlysuitedforpatientmanagement[a].Weproposeacompleteofferwithmorethan15dyescoveringallqPCRchannels.OurMGB-Probescanbedeliveredin6,20and50nmol,driedorinsolution(TEorH2O).Formaximalconvenience,a10nmoldriedaliquotedformatisalsoavailableforthe20and50nmolquantities,atnoadditionalcost.TheprobesareavailableinIVDgrade,uponrequest.n

LC Hybridization probes

MGB Probes [a, b]

*Please note that due to the use of various scales some discrepancies

might be observed between the right and left parts of this figure. Only available in FRET quenching. QR:

indicative quenching range.

a. Please note that MGB probes from Eurogentec are intended for human patient management use only.b. Eurogentec is an authorized supplier of MGB Probes for MGB-licensed IVD kit manufacturers. No restriction applies with respect to the field of use in this case.

Fluorescent DyesCascade blueAlexa Fluor 405®

AMCA-XAlexa Fluor 350®

Pacifi c Blue™

Marina blue®

ATTO 390ATTO 425EDANSCy®2ATTO 465BODIPY® 493/503BODIPY® FL-X/ BODIPY® FLDY-475XLAlexa Fluor® 488FAMOregon Green® 500Hilyte Fluor™ 488Oregon Green® 488ATTO 488FluoresceinOregon Green® 488-XOregon Green® 514ATTO 495Oyster-500DY-495-05DY-495-X5Rhodamine Green™-XDY-505-05DY-505-X5Lucifer YellowRhodamine Green™

TETAlexa Fluor® 430BODIPY® 507/545CAL Fluor Gold 540ATTO 520JOEYakima Yellow®

BODIPY® R6GBODIPY® 530/550ATTO 532VICAlexa Fluor® 532DY-500XLHEXCAL Fluor™ Orange 560 DY-485XLCy®3Alexa Fluor® 555Hilyte Fluor™ 555Quasar 570DY-554BODIPY® 558/568BODIPY® 564/570BODIPY® R-XOyster-556DY-555DY-556Alexa Fluor® 546DY-547NEDDragonfl y orange™

ATTO 550DY-560Rhodamine Red™-XTAMRADY-510XLBODIPY® 581/591CAL Fluor® Red 590ATTO 565PETFAR-FuschiaDY-590ROXAlexa Fluor 568Texas Red®-XCY®3.5Hilyte Fluor™ 594CAL Fluor™ Red 610BODIPY® TR-XAlexa Fluor® 594ATTO 590ATTO 594Alexa Fluor® 610DY-610DY-480XLATTO 610DY-600XLCAL Fluor® Red 635BODIPY® 630/650LC Red 640DY-615ATTO 620Alexa Fluor® 633PULSAR 650LIZDY-630DY-632DY-633ATTO 633DY-631ATTO 635ATTO 637Cy®5DY-520-XLAlexa Fluor® 647BODIPY® 650/665Oyster-645Hilyte Fluor™ 647Quasar™ 670ATTO 647ATTO 647NWellRed D4-PADY-635DY-636DY-647DY-650Oyster-656FAR-BlueDY-651ATTO 655Alexa Fluor® 660Hilyte Fluor™ 680DY-673DY-675DY-676ATTO 680Alexa Fluor® 680LC Red 705WellRED D3-PACy®5.5DY-681DY-680IRD 700DY-690ATTO 700Alexa Fluor® 700DY-700DY-701ATTO 725DY-730DY-732DY-731ATTO 740Cy®7Hilyte Fluor™ 750WellRED D2-PAAlexa Fluor® 750DY-750DY-751FAR-Green TwoDY-781DY-782DY-776DY-780IRD 800IRIS-Green One

Max Abs (nm)396401353346416362390436336489453500504493495494499495499501492495506495500503503503505505428505521433508522525520530530534532538532505535538485546555548553549559564544556547548556557546554554559560556509581569563558567580575578583588590596588590594601612609500615603618625625621619632460638636637637629637635635646520650650645647652645644650647645653653656660653663663673678674674680679685685683691690685691700702707706725732736736740743750754749747751772783782771770795800

Max Em (nm)410421442442451459479484490506508509510514519520519521523523525525526527527528528528530530532535536541543544545548549550551553554554555556559560563565566568568568569570570572573573574575576576578580588590591591592595597599602603603604610617616617624627628629630634634637640640641643650650655657657657657658659659662664665665666667669669669670671671672674674678678684690696699699699700702705706707708709710714719723730731752758759760764767770778775776779788800800801810819820

IR

UV

Emission color

DABCYLlabs(max): 479 nmQR: 390- 510 nm

BHQ-1®

labs(max): 534 nmQR: 480- 580 nm

BHQ-2®

labs(max): 579 nmQR: 550-650 nm

BHQ-3®

labs(max): 680 nmQR: 620-730 nm

Deep Dark Quencher Ilabs(max): 440 nmQR: 380-500 nm

Eclipse® Dark Quencherlabs(max): 522 nmQR: 460-570 nm

Deep Dark Quencher IIlabs(max): 595-650 nmQR: 570-680 nm

Fluorescent dyes

Excitation Emission

Primer

Primer

Transfer Probe5'

3'5' 5'

F1 F2

Primer

Primer

5'

3'

5'5'

F Q

MGB

Excitation Emission

Primer

Primer

Transfer Probe5'

3'5' 5'

F1 F2

Primer

Primer

5'

3'

5'5'

F Q

MGB

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Plexor™ Primers

ThePlexor™primertechnologyisatechniqueforReal-TimeqPCRadaptedtomultiplexassays.Thistechnology

takesadvantageofthehighlyspecificinteractionbetweentwomodifiednucleotides.Thesetwonovelbases,calledisoguanine(iso-dG)and5-methylisocytosine(iso-dC),formauniquebasepairwhenincorporatedindouble-strandedDNAandpaironlywitheachother.InPlexor™reactions,onePCRprimerissynthesizedwithaniso-dCresidueandafluorescentlabelatthe5’-end.ThesecondPCRprimerisastandardunlabelledoligonucleotide.n

Adapted for Real-

Time qPCR Multiplex

Assays

Simplifyk ISO 15189 Accreditationk Reagents QCk Set Up

Improvek Quality Standardsk Cost Efficiency vsk Assay Precision

Assay Dispensing Service

“

“

A happy Customer Laboratory of Microbiology London Hospital, UK

I have about five lots of 12 strips to get through and there has been no diminution in CT value even though they expired...My colleagues have been looking on in envy and are keen to experience the joys of pre-aliquoted strips. Recently we had a meeting to discuss what we might need if we used pre-aliquoted strips for all our current routine qPCR assays.

Dr. A. FerroniLaboratory of Bacteriology APHP Necker Hospital, France

Saves a lot of technician handling time and reduces 16S contamination during MasterMix preparation.

Pr. Rémi CHARREL Laboratory of Bacteriology APHM Timone Hospital, France

The best solution to sim-plify accreditation of in-house PCR & qPCR as-says, combining optimal profitability, efficiency and ease-of-use.

Secure your in-house qPCR assays

More info: www.eurogentec.com/dispensing.html h [email protected]

Testimonials

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PrincipleofsiRNA-mediatedRNAinterference

TT

TT

TT

TT

3' 5'

siRNA

m RNA

RISC form ation

Target recognition

Recycling

Target degradation

RISC

RNAi Oligonucleotides


•1000nmol*Backbone RNA,LNA®,2’O-MeRNA andalllinkagesModifications 5’:Phosphate,6-FAM,Cy®3,Cy5®,

TET,HEX,...3’:DABCYL,TEG-Cholesteryl,TAMRA...

Purifications SePOPDesalting,RP-HPLCorPAGEQuality Control MALDI-TOFMSFormat DriedPackaging 2mLtubeDocumentation TechnicalDataSheetsiRNA Design FreeandguaranteedShipping AtRoomTemperature


RNA INTERFERENCE isamechanismofgenesilencingatthemRNAlevel.Thisphenomenonistriggeredbysmallinterfering(si)RNAsandmicro(mi)RNAs.siRNAsandmiRNAsarecapableofinhibitinggeneexpressionbyeitherdirectingthedegradationofhomologousmRNAtargetsorinducingtherepressionoftranslationofmRNAtargetswhichhaveincompletecomplementarity.n

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Custom siRNA Duplexes

BycompilingthemostdocumentedstrategiesforthedesignofeffectivesiRNAsequences,Eurogentechasco-developedanexclusivesiRNAdesignplatform.PhD-levelscientistsofourdesignteamusethisreliableinterfaceto

designcustomsiRNAforanytargetofyourchoice.n

k Positive controlsconsistofsiRNAdirectedagainstarangeofendogenousandreportergenes.Theyareavailablein5,10and20nmolfinalquantities.Eachcontrolcontains1siRNAduplex.AllsiRNAcontrolduplexesarePAGEpurifiedand100%MALDI-TOFMassSpectrometrycontrolled.Thesequencesproposedarevalidatedandpublished.n

Clear-MiR™ miRNA Inhibitors

Clear-MiR™miRNAinhibitorsarechemicallymodifiedantisenseRNAOligonucleotidesoptimizedto

specificallytargetmiRNAmoleculesincells.n

Add-MiR™ miRNA Mimics

Add-MiR™OligonucleotidesareCustomdouble-strandedsyntheticmiRNAmimickingtheactionofendogenousmiRNAs.n

Control siRNA Duplexes

InordertomonitoryoursiRNAexperimentconditions,EurogentecprovidessiRNAcontrolduplexesandkitsincludingnegativeandpositivecontrolsnecessarytovalidyourexperiment.

k Negative control isconstitutedofsiRNApresentingnohomologywithanyknowneukaryoticgene.siRNAcontrolisalreadyannealedandshippedinsolution.Theproposedsequenceisproperlyvalidated.

siRNA miRNA

The easiest and most

efficient way to achieve

RNAi

The best way to target

specific messenger

RNA

>Note• The antisense strand must either have a free

5’-OH (by default) or 5’-phosphate terminus. • Certain modifications can sometimes be

useful e.g. to increase stability or cellular uptake. Modifying siRNA with cholesterol is used to facilitate tissue / cellular uptake.

• Various fluorescent dyes can be coupled to the 5’-end of the sense strand oligonucleotide to track transfection efficiency of the corresponding duplex.

>Note Clear- MiR™ miRNA Inhibitors and Add-MiR™ miRNA Mimics are available with different labels and can be linked to cholesterol to increase cellular uptake. On request, peptides can also be covalently linked.

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Universal primersName Sequence Bases Tm (°C)

16S rRNA For AGA GTT TGA TCC TGG CTC AG 20 55.2

16S rRNA Rev ACG GCT ACC TTG TTA CGA CTT 21 57.4

3’ RACE PCR GGC CAC GCG TCG ACT AGT AC 20 60.6

Anchored Oligo dT (20) TTT TTT TTT TTT TTT TTT TV 20 39.2

Anchored Oligo dT (22) TTT TTT TTT TTT TTT TTT TTV N 22 42.8

Bluescript KS TCG AGG TCG ACG GTA TC 17 53.3

Bluescript SK CGC TCT AGA ACT AGT GGA TC 20 52.4

cDNA Cloning Primer GGC CAC GCG TCG ACT AGT ACT TTT TTT TTT TTT TTT TV 38 64.8

EGFP-C CAT GGT CCT GCT GGA GTT CGT G 22 61.2

EGFP-N CGT CGC CGT CCA GCT CGA CCA G 22 67.2

G3PDH For ACC ACA GTC CAT GCC ATC AC 20 58.6

G3PDH Rev TCC ACC ACC CTG TTG CTG TA 20 59.7

M13 Forward (-20) GTA AAA CGA CGG CCA GT 17 53.0

M13 Forward (-41) CGC CAG GGT TTT CCC AGT CAC GAC 24 65.5

M13 Reverse (-27) CAG GAA ACA GCT ATG AC 17 47.3

M13 Reverse (-48) AGC GGA TAA CAA TTT CAC ACA GG 23 57.2

Neomycin For CTT GGG TGG AGA GGC TAT TC 20 55.6

Neomycin Rev AGG TGA GAT GAC AGG AGA TC 20 54.0

Oligo dT, 15mer TTT TTT TTT TTT TTT 15 29.7

Oligo dT, 16mer TTT TTT TTT TTT TTT T 16 32.1

Oligo dT, 18mer TTT TTT TTT TTT TTT TTT 18 36.0

Oligo dT, 20mer TTT TTT TTT TTT TTT TTT TT 20 39.1

PCMV Forward CGC AAA TGG GCG GTA GGC GTG 21 64.8

pET 3’ CTA GTT ATT GCT CAG CGG 18 50.6

pET 5’ (T7) TAA TAC GAC TCA CTA TAG G 19 45.3

pET Upstream ATG CGT CCG GCG TAG A 16 56.7

pGEX 3’ CCG GGA GCT GCA TGT GTC AGA GG 23 65.2

pGEX 5’ GGG CTG GCA AGC CAC GTT TGG TG 23 67.0

ROSA26 Promoter For AAA GTC GCT CTG AGT TGT TAT 21 53.2

ROSA26 Promoter Rev GGA GCG GGA GAA ATG GAT ATG 21 56.3

SP6 Promoter TAC GAT TTA GGT GAC ACT ATA G 22 50.0

SP6 Upstream ATT TAG GTG ACA CTA TAG 18 42.8

T3 Promoter AAT TAA CCC TCA CTA AAG GG 20 50.4

T7 Promoter TAA TAC GAC TCA CTA TAG GG 20 48.3

T7 Terminator GCT AGT TAT TGC TCA GCG G 19 54.1

Universal Primers

Final Yield 1OD/5nmolBackbone DNAModifications NonePurifications RP-HPLCQuality Control MALDI-TOFMS+CGEFormat DriedPackaging 2mLtubeDocumentation TechnicalDataSheetShipping AtRoomTemperature

DURING SEQUENCING,primersareannealedtothedenaturedDNAtemplatetoprovideaninitiationsitefortheelongationofthenewDNAmolecule.PrimerscaneitherbespecifictoaparticularDNAnucleotidesequenceortheycanbe“Universal.”

NucleicacidsequenceanalysisisanextensivelyappliedmethodinGenomicsstudies.UniversalprimersarecomplementarytonucleotidesequencesthatareverycommoninaparticularsetofDNAmoleculesandcloningvectors.Thus,theyareabletobindtoawidevarietyofDNAtemplates.

AllUniversalOligonucleotides(seetablejustbeside)areprovidedRP-HPLCpurified,MALDI-TOFMSandCGEcontrolled.Theyaresentdriedin2mLtubewithatypicalfinalyieldof1OD/5nmolnextdayafterorderreceipt(subjecttoin-stockavailability).n

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PNA FISH Probes

Length 18basesSynthesis scale 5nmolBackbone PNAModifications FAM,Cy®3,FITC,Cy5®,TMRPurifications RP-HPLCQuality Control MALDI-TOFMS+RP-HPLCFormat DriedPackaging 2mLtubeDocumentation TechnicalDataSheetShipping Atroomtemperature

IN PRINCIPLE, FLUORESCENCEinsituhybridization(FISH)shouldbeabletoprovideinformationonthetelomerelengthofindividualchromosomes.Directlylabelledoligonucleotideprobesareattractivecandidatesforsuchanalysisbecausetheyaresmall(goodpenetrationproperties),singlestranded(nodenaturationofprobe)andtheirsynthesisiscontrolled.HowevertheefficiencyofoligonucleotidehybridizationsfortelomericrepeatshasnotbeensufficienttoextendthisapproachbeyondqualitativestudiesofTTAGGGrepetitionsinchromosomesofvariousspecies.Recently,itwasshownthatpeptidenucleicacid(PNA)oligonucleotideprobeswillhybridizewithcomplementaryoligonucleotidesequencesandthattheresultingduplexesaremorestablethanDNA/DNAorDNA/RNAduplexes.InPNA,thechargedphosphate-(deoxy)ribosebackboneofconventionalDNAandRNAoligonucleotidesisreplacedbyanunchargedbackboneofrepeatingN-(2-aminoethyl)-glycineunitslinkedbypeptidebonds.IncomparisonwithDNAoligonucleotides,PNAoligomersdemonstrateahighersequencespecificity,animprovedstability,abetterreproducibility,andalowerbackground.DuetothehigherTmofPNA/DNAduplexes,short(18-mer)telomerePNA(CCCTAA)3arewidelyused.n

s TelC-FAMs TelC-Cy3s TelC-Cy5s TelC-FITCs TelC-TMRs TelG-FAMs TelG-Cy3s TelG-Cy5s TelG-FITCs TelG-TMR

>Note PNA FISH Probes are also available to detect chromosome aberrations in the centromer. Please contact us at [email protected] for more information.

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The Unique solution you deserve!

Unique Oligonucleotides

Becauseyourexperimentsrequirealwaysmorecustomisation,UniqueOligonucleotidesbringyoutheperfectsolutionwhenyouneedoligoswithatypicalchemistries,raremodifications,customsynthesisscaleorfinalguaranteedamount,personalizedlabelsandpackagings…n

WITH OVER 25 YEARS EXPERIENCEinOligonucleotides,Eurogentechassynthesizedsophisticatedchimeras,siRNAcoupledtovariouspeptides,highlycomplexsequences,veryshortlabelledOligonucleotides,G-stretches,aptamers…inguaranteedquantitiesfromnanogramstomilligrams.

Ifyoudonotfindtheoligonucleotidethatbestsuitsyourneeds,thinkaboutuniqueoligosandcontactusatunique@eurogentec.commentionningthespecificationsofyourUniqueOligonucleotides(sequenceorlength,chemistries,modifications,purifications,expectedpurity,synthesisscaleorfinalamount,format,packaging…).Youwillreceivethecorrespondinginformationintermsoftechnicalfeasibility,pricingandturnaroundtimeswithin2workingdays.n

SpecificationsLength From2to225basesSynthesis scale CustomizedBackbone UsualoratypicalChemistryModifications CommonorraremodificationsPurifications SePOPDesalting,

RP-Cartridge•Gold™,HPLC,PAGE, DualHPLC,UltraPureGold™

Quality Control AdaptedtoyourneedsFormat AdaptedtoyourneedsPackaging AdaptedtoyourneedsDocumentation TechnicalDataSheet

CustomdocumentationShipping AtRoomTemperature

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THERE IS OFTEN SOME CONFUSION surroundingtheissuesofsynthesisscaleandminimumguaranteedyieldinOligonucleotidemanufacturing:

k ThesynthesisscalereferstotheamountofrawmaterialusedtostartthesynthesisofOligonucleotides.

k Theyieldcorrespondstotheamountoffinalproductrecoveredattheendofthesynthesisandpurificationprocesses.

Thelength,thesequence,thetype/numberofmodificationsandthepurification,stronglyinfluencetheamountofsynthesisedmaterial,andtheproportionoffull-lengthOligonucleotides.Basedonthat,Eurogentecdefinedaminimumguaranteedyieldinnmolesforallproductcategories(seetablebeside).

Inmanycasestheminimumguaranteedyieldsrepresentonlyareferencebecausethedeliveredquantitiesaresignificantlyhigher. n

Minimum Guaranteed Yields

Minimum guaranteed yields in nmolSynthesis scale 10 40 200 1000 2500 5000 10000 20000

Purification

SePO

P

RP-C

artri

dge-

Gold

™

HPLC

(RP

or IE

X)

SePO

P

RP-C

artri

dge•

Gold

™

HPLC

(RP

or IE

X)

PAGE

(3)

Dual

HPL

C

SePO

P

RP-C

artri

dge•

Gold

™

HPLC

(RP

or IE

X)

PAGE

(3)

Dual

HPL

C

Ultra

Pure

Gold

™

SePO

P

RP-C

artri

dge•

Gold

™

HPLC

(RP

or IE

X)

PAGE

(3)

Dual

HPL

C

Ultra

Pure

Gold

™

SePO

P

HPLC

(RP

or IE

X)

Dual

HPL

C

SePO

P

HPLC

(RP

or IE

X)

Dual

HPL

C

SePO

P

HPLC

(RP

or IE

X)

Dual

HPL

C

SePO

P

HPLC

(RP

or IE

X)

Dual

HPL

C

Range Product Length

Cust

om O

ligon

ucle

otid

es

Non-Modified(DNA only)

5-9 - - - - - - - - 60 50 30 20 15 - 180 100 80 40 40 - 450 200 100 900 400 200 1800 800 400 - - -10-19 - - - - - - - 3 70 60 45 30 23 15 200 140 100 70 50 30 500 250 125 1000 500 250 2000 1000 500 4200 2100 105020-39 5 4 - 20 16 10 4 2 60 50 30 20 15 - 190 120 90 40 45 - 475 225 115 1000 500 250 2000 1000 500 4200 2100 105040-59 3 2 - 10 8 5 2 1 30 25 15 12 7 - 115 60 45 20 20 - 285 110 55 600 230 115 1200 460 230 2500 1000 50060-79 - - - - 6 - 2 - - 18 - 8 - - - 40 - 14 - - - - - - - - - - - - - -80-99 - - - - - - 1 - - - - 3 - - - - - 5 - - - - - - - - - - - - - -

100-139 - - - - - - - - - - - 2 - - - - - 3 - - - - - - - - - - - - - -

Modified (1)

(including DNA, RNA, 2’ O-Me RNA & LNA®)

5-9 - - - - - - - - - - 12 - - - - - 25 - 12 - - 60 30 - 125 60 - 250 125 - - -10-19 - - - 12 6 5 4 1 35 20 17 15 8 - 70 40 35 30 15 - - 90 45 - 190 95 - 380 190 - - -20-59 - - - 8 5 4 3 1 20 15 12 10 6 - 45 35 25 20 12 - - 65 30 - 135 65 - 275 130 - 600 27560-139 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -


Double-Dye probes (2) 8-38 - - <2(4) - - 4 - - - - 12 - - - - - 25 - - - - 65 - - 135 - - 275 - - 600 -Molecular Beacons 32-50 - - - - - - - - 4 - - - - - 12 - - - - 30 - - 65 - - 130 - - 275 -

MGB Taqman Probes 8-30 Delivered quantity: 6, 20 or 50 nmol

RNAi OligonucleotidessiRNA Duplexes Non-Modified (5) 21-27 5 - 1 15 - 8 3 - 40 - 25 13 - - 160 - 70 25 - - - 175 - - 360 - - 720 - - 1500 -

siRNA Duplexes Modified (1) 21-27 - - - 8 5 4 3 1 20 15 12 10 6 - 45 35 25 20 12 - - 65 30 - 135 65 - 275 130 - - -

NGS OligonucleotidesRP-Cartridge purified

20-85 Minimum delivered quantity: 10 nmolRP-HPLC purified

Universal Primers - 15-38 - - - - - 5 - - - - - - - - - - - - - - - - - - - - - - - - - -

Unique Oligonucleotides - 2-225 On Request - please contact us at [email protected]

Table: (1) Between 5 and 59 bases length single-modified Oligonucleotides. Eurogentec does not provide minimum guaranteed yield for modified Oligonucleotides greater than 59 bases. Post-synthesis modifications (not compatible with SePOP and RP-Cartridge•GoldTM purification) may yield 50% less than the above stated values. A lower yield may result from poly-modifications and/or strong secondary structures ; (2) Double-Dye probes only result from the combination of a 5’ fluorescent dye and a 3’ quencher. Postsynthesis modifications may yield 50% less than the above stated values. (3) Except for Oligonucleotides with GC-rich regions; (4) Only available for Double-Dye FAM-TAMRA 10 nmol and FAM-BHQ1® 10 nmol ; (5) Non-modified siRNA’s only include 3’ dTdT overhang. For more information, please contact our Oligo Centre at [email protected] or visit our website www.eurogentec.com.

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Shipping methods

Eco-Logik DeliverykBylocalMailtoreducetheglobalecologicalOligonucleotidesinyourmailbox.AvailableforBelgium,FranceandMonaco

Express DeliverykAll OligonucleotideskBy Express courrierToreceiveyourOligonucleotidesasfastaspossible(24to48hours)inyourhands.kSame day shipping option-Forordersreceived

before10.00AM(CentralEuropeanTime)

-ForcustomOligonucleotides(max24),10/40nmolscale,5-30DNAbases,unmodified,SePOPdesaltedorRP-Cartridgepurified.

Selecttheshippingmethodthatbestsuitsyourneeds.YoucanchoosetheEco-LogikDeliveryortheExpressDelivery(withapossible"SamedayshippingOption").ThedeliverytimewillvarydependingonthespecificationsofyourOligonucleotides(seetablebeside)

Shipping

>Note The online tracking allows you to check the statements of your oligonucleotide orders at any time.

Delivery times (in working day)

Between 1 and 14 Oligonucleotides Purifications

Range Product Length

SePO

P

RP-C

artri

dge•

Gold

™

HPLC

(RP

or IE

X)

PAGE

Dual

HPL

C

Ultra

Pure

Gold

™


Non-Modified (DNA Only)

5-9 - 4-5 5 6 7 7

10-39 2-3 4-5 5 6 7 7

40-59 5 6 7 8 - 9

60-79 - 6 - 8 - 9

80-139 - - - 10 - 11

Modified(including DNA, RNA, 2’ O-Me RNA & LNA®)

10-39 5 7 7 8 9 9

40-59 7-8 9-10 9-10 10-11 11-12 11-12


Double-Dye Probes 8-38 - - 7 - - -

Molecular Beacons 32-50 - - 12-15 - - -

MGB Taqman Probes 8-30 - - 5-7 - - -

RNAi Oligonucleotides siRNADuplexes 21-27 5-7 - 9-10 10-11 - -

NGS Oligonucleotides - 20-85 - 4-6 5-7 - - -

Universal Primers - 15-38 - - 2-3 - - -

Unique Oligonucleotides - 2-225 On Request

For more Oligonucleotides or Unique Oligonucleotides, please feel free to contact us at [email protected] to receive more details in terms of delivery schedules. 5'AP, BSA, HRP or SBP Conjugation : 3-5 WD Extra. Additional Purification or Services: 2 WD Extra ; Fax Ordering: 1 WD Extra

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DocumentationEACH Oligonucleotideisprovidedwithatechnicaldatasheet.Otherdocumentationscouldbeaddeddependingontheoligonucleotidetype.Allthedocumentsaresentaspdffilestoyourshippingemailaddress.n

OLIGONUCLEOTIDETechnical Data Sheet (TDS)

YOUR ORDER NUMBER NRU / SON DATE

1111111110 *1111111110**1111111110**1111111110**1111111110* 28-Oct-10 CUSTOMER ADDRESS / INSTITUTION Kerdraon B. BELGIUM 4000 Liège OLIGO NAME BATCH SCALE CHEMISTRY BASES Function control 2500006 200 nmol DNA 50 SEQUENCE (5ʼ 3ʼ) * Phosphorothioate linkage1 GTA-CK1-A2T-TMT-AN3-CT4-56A-WCS-GHT-Y7A-TAC-8CG-ATA-9CG-TCU-UTT-GC

BACKBONE (BASES + LINKAGES)A 8 GC (%)1 26.0 Quantity (OD@260nm) 12.56C 8 MW (g.mol-1)1 16724.5 (nmol) 25.2G 5 Ext. coeff. (l.mol-1.cm-1)1 498800 (µg) 421U / T 2 / 11 Tm (°C) 1 59.5Others 9 Wobbles 7 1 Theoretical values MODIFICATION(S)Mod 3ʼ BHQ-1 ™ 554.49 Mod 4 LNA T ® 332.2 1Mod 5ʼ Atto 565 672.74 Mod 5 2' OMe C 319.21 1

Mod 6 2' OMe G 359.2 1Mod 1 LNA A ® 341.21 1 Mod 7 2' OMe U 320.2 1Mod 2 LNA C ® 331.22 1 Mod 8 5-Me dC 303.21 1Mod 3 LNA G ® 357.21 1 Mod 9 Carboxy dT 360.22 1PURIFICATION(S) HPLC-RP HPLC-IEX ADDITIONAL SERVICE(S) FORMAT Dried ALIQUOTING

Number of Aliquots 3

QUALITY CONTROL(S) MS HPLC-RP CGE PassedDELIVERY CONDITION Room Temperature RECONSTITUTION To Make up to 100 µM concentration add 252 µl of recommended buffer. To Make up to 20 µM concentration add 1.26 ml of recommended buffer. To Make up to 5 µM concentration add 5.04 ml of recommended buffer. STORAGE 4 °C as dried, -20 °C as solution COMMENTS General: Tm % GC calculated with 50 mM NaCl and 0 % formamide (PCR conditions) Specific: HELP EUROPE NORTH AMERICA

If you have any questions feel free to call our Oligo Centre depending on your location: [Belgium, France, Germany, Luxembourg, The Netherlands, United Kingdom, Switzerland,

Austria] 00 800 666 00 123 (European toll free number), or send an email to [email protected]

If you have questions, please contact our Technical Support Department at +1 (858)793-2661 or +1 (858) 793-6436 or send an email to

[email protected]

Eurogentec S.A. Liège science park- Rue Bois Saint-Jean 5 - 4102 SERAING BELGIUM Tel : +32(0)4 372 74 00 - Fax : +32(0)4 264 07 88 E-mail : [email protected]

RPM Liège T.V.A.-(BE)-0427.348.346;IBAN : BE02 2400 7776 8540 BIC : GEBABEBB Fortis Bank 240-0777685-40 www.eurogentec.com

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RoutineMALDI-TOFMassSpectrometry(MS)analysisofastandard15RNAbasessequence.The15RNAbaseswasnotpurifiedpriortoanalysis.

TDS:TechnicalDataSheet;MS:MassSpectrometry;HPLC:HighPerformanceLiquidChromatography;CGE:CapillaryGelElectrophoresis.(1)Alwaysprovidedupto60baseslongOligonucleotides.(2)Ifapplicable.(3)IEX-HPLCforshortOligonucleotides.(4)ExceptforSePOPdesaltedOligonucleotides.(5)Optional.Fortechnicalreasonsthisgeneralrulemaybeadaptedtoprovideyouwiththemostsuitableandusefuldocumentation.

Sample tube Technical Data Sheet (TDS)Documentation

TDS MS(1) HPLC(2) CGE(3)


Unmodified

Modified ✓(4)

UltraPureGold™ ✓ ✓


RNAi Oligonucleotides siRNA Duplexes (4)

Universal Primers ✓

Unique Oligonucleotides (5) ✓(5)

Sample tube labela), Pag e \

GTA-CK1-A2T-TMT-AN3-CT4-56A-WCS-GHT-Y7A-TAC-8CG-ATA-9CG-TCU-UTT-GCWg/molµg nmol°C EU

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TEC

C U S T O M O L I G O

A

A

BC

D

E

BCDE

A Nameofyouroligonucleotide

B Sequence

C Molecularweight

D Quantities(µg,nmole,OD)

E Modifications

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A Nameofyouroligonucleotide

B Sequence

C Molecularweight

D Quantities(µg,nmole,OD)

E Modifications

Ordering information

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Oligonucleotides

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# Storage and stability

Tips andtricks

# Oligonucleotide quantification1OD260(OpticalDensity)unitisdefinedastheamountofoligonucleotidewhich,whendissolvedinavolumeof1.0mL,resultsinanabsorbanceof1.0whenmeasuredat260nmina1cmpath-lengthquartzcuvette.1OD260unitcorrespondstoapproximately33µgofsinglestrandDNA.Theserelationships,however,canbeinaccurateforshortfragmentsofDNA,suchasOligonucleotides.Basecompositionandevenlinearsequencewillaffectopticalabsorbance.HencetheprecisevalueoftheODtomassrelationshipisuniqueforeacholigonucleotide.

Example: 1.0 OD260 of CCCCCCCCCC (10 bases) equals 39 µg whereas 1.0 OD260 of AAAAAAAAAA (10 bases) equals only 20 µg.

WecarefullymeasuretheODvalueforyourCustomOligonucleotidebymeasuringtheabsorptionat260nmusingUVspectrophotometer.ThisinformationisprovidedontheoligonucleotideTechnicalDataSheetasthenumberofOD260units.TheamountofoligonucleotideexpressedinnanomolesandmicrogramsisderivedfromtheODmeasurement.

> To quantify your Oligonucleotides, make an aliquot of the resuspended Oligonucleotides to a final volume of 1 mL of dH2O and vortex for a few seconds. Measure the absorbance of this dilution at 260 nm (A260). Use the formula below to calculate the concentration of Oligonucleotides in your stock solution.

This formula is valid for an absorption of A260 ≤1.2.

Calculate the number of nanomoles present given an OD reading and extinction coefficient: Nanomoles = ( OD260 / ε260) × 106

Example: 1 OD260 unit of primer M13 Forward, 5’-GTA AAA CGA CGG CCA GTG-3’ Molar extinction coefficient (ε260) = 182.800 L / (mole x cm) Nanomoles = (1.0 / 182.800 ) × 106 = 5.47 nmoles

Convert the amount in nanomoles to micrograms:Micrograms = Molecular Weight × Nanomoles × 10-3

Example: 1 OD260 unit of primer M13 Forward, 5’-GTA AAA CGA CGG CCA GTG-3’ Molecular Weight = 5558.7 Micrograms = 5558.7 × 5.47 × 10-3 = 30.4 µg

equation

Concentration in µg/mL = A260 × dilution factor x Weight per OD of stock solution (in µg / OD)

# Reconstitution

k Spinthetubebrieflytocollectthepelletinthebottomofthetube.k Addanappropriatevolumeofrecommendedbuffer.k Allowthetubetostandafewminutes.k Vortexthetubefor15secondes.k Refertothededicatedtechnicaldatasheetformoreinformation.

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# Calculation of the molecular weight :

Anhydrous MW (g/mol) = εIndividual Base MW + εIndividual Mods MW - 63.98 + 2.016

# Calculation of the molar extinction coefficient

where∑NearestNeighbouristhenearestneighbourconstantforapairofbases,∑Individualistheconstantforanindividualbase,andnisthelengthoftheoligonucleotide.

> Note ∑Modificationis not known for all modifications.

For DNA bases:MWdA=313.21;MWdC=289.18;MWdG=329.21;MWdT=304.20;MWdU=290.17;MWdI=314.19

For RNA bases:MWDNAcounterpart+16.WhendeterminingtheweightofUracil(rU)startwithdUandnotdT

For LNA bases:MWdA=313.21;MWdC=289.18;MWdG=329.21;MWdT=304.20;MWdU=290.17;MWdI=314.19

For 2’ O-Methyl bases:MWDNAcounterpart+30.03.WhendeterminingtheweightofmUstartwithdUandnotdT

For phosphorothioated bases:MWDNAcounterpart+16.06

# Mixed basesMixedbases(alsoknownasdegenerateorwobblebases)followtheIUBcodes:D=A/G/TM=A/CH=A/C/TW=A/TR=A/GY=C/TV=A/C/GS=C/GK=G/TN=A/G/C/TB=C/G/T

Oligonucleotidesynthesisedwithmixedbasesgivesafinalproductthatisaheterogeneouspopulationofdistinctspecies.MW,Tmandextinctioncoefficientmaybestronglyaffectedbymixedbaseaddition. Ratherthanreportingthevariousvaluesforeachcomponent,asinglevalueisgiven.

# ChemistriesACGTA=DNA(ACGUA)=RNA[ACGUA]=2'O-MeRNA{ACGTA}=LNA®

A*C(G*U*)A=Phosphorothioates

# Oligonucleotide scale determination (PCR and Real Time qPCR)

Number of PCR reactions Quantity of DNA (µg) OD260

Synthesis scale (nmol)

50-500 33 1 10

200-2000 132 4 40

600-6000 396 12 200

1000-10000 660 20 1000

Table1.OligonucleotidescaledeterminationfordifferentnumberofPCRreactions(Finalvolume:100µl,FinalconcentrationofOligonucleotides:0.5µM,Oligonucleotidelength:20bases).

Number of PCR reactions Quantity of DNA (µg) OD260

Synthesis scale (nmol)

300-600 15 0.5 10

600-1200 30 1 40

2000-4000 90 3 200

4000-8000 200 6 1000

Table2.OligonucleotidescaledeterminationfordifferentnumberofReal-TimeqPCRreactions(Finalvolume:50or25µl,FinalconcentrationofOligonucleotides:100nM,Oligonucleotidelength:20bases).

# Genetic code5’OH U C A G 3’OH

U

UUU Phe UCU

Ser

UAU Tyr UGU Cys UUUC UCC UAC UGC CUUA Leu UCA UAA Stop UGA Stop AUUG UCG UAG Stop UGG Trp G

CCUU

Leu

CCU

Pro

CAU His CGU

Arg

UCUC CCC CAC CGC CCUA CCA CAA Gln CGA ACUG CCG CAG CGG G

A

AUUIle

ACU

Thr

AAUAsn

AGUSer U

AUC ACC AAC AGC CAUA ACA AAA

LysAGA

Arg AAUG Met ACG AAG AGG G

G

GUU

Val

GCU

Ala

GAUAsp

GGU

Gly

UGUC GCC GAC GGC CGUA GCA GAA Glu GGA AGUG* GCG GAG GGG G

Table3.TheGeneticCodeandits64combinations.The3stopcodonsarecalledOchre=UAA,Amber=UAGandOpal=UGG.*GUGcodesforMetifintheinitiatorposition.

n-1 n-1 n

121

ε260 = 2× (ε ε ) - ε ε + ε ε Nearest Neighbour

Individual Modification

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Alexa Fluor®isaregisteredtrademarkofMolecularProbes,Inc.(awhollyownedsubsidiaryofInvitrogenCorp.)

BHQ-0®, BHQ-1®, BHQ-2®, BHQ-3®areregisteredtrademarksofBiosearchTechnologies,Inc.

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License statements

1. LicenseddsDNA-BindingDyeResearchKits

k UseofthisproductiscoveredbyoneormoreofthefollowingUSpatents

andcorrespondingpatentclaimsoutsidetheUS:5,994,056,and6,171,785.

Thepurchaseofthisproductincludesalimited,non-transferableimmunity

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expressly,byimplication,orbyestoppel.Thisproductisforresearchuse

only. Diagnostic uses under Roche patents require a separate license

fromRoche.Furtherinformationonpurchasinglicensesmaybeobtained

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CentreDrive,FosterCity,California94404,USA.

2. Licensed5’NucleaseResearchKits

k Use of this product is covered by one or more of the following US

patents and corresponding patent claims outside the US: 5,804,375,

5,538,848,5,723,591,5,876,930,6,030,787and6,258,569.Thepurchase

of this product includes a limited, non-transferable immunity from

suit under the foregoing patent claims for using only this amount of

productforthepurchaser’sowninternalresearch,Norightunderany

otherpatentclaimandnorighttoperformcommercialservicesofany

kind, includingwithoutlimitationreportingtheresultsofpurchaser’s

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use only. Diagnostic uses under Roche patents require a separate

license fromRoche.Further informationonpurchasing licensesmay

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foreignequivalentsfromAlnylamPharmaceuticals,Inc.,Cambridge,USA

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k LNA® Oligonucleotides produced under license from Exiqon A/S

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9. C-5PropyneandG-clampmodifiedOligonucleotides

k C-5 Propyne and G-clamp modified oligonucleotides are covered

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ownedbyAnaSpec,Inc.:USP7,465,810,USP7,754,893,USP7,820,783,USP

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