1 introductionn

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Welcome to MCDB 306

Genetics Laboratory

Nhu Do


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Lecture 01 January 11th

• Todays !utline – Course "olicies

– #ntroduction to"ro$ects andDrosophilamelanogaster

• %nnouncements – #& you are 'ait listed

&or a lab( "lease )o tothe lab you 'ant toattend and see i&there is s"ace

– We 'ill &ollo' 'ait listorder to *ll anyremainin) s"ots

Wee+ o& January 1,th and 'ee+

!& January -.th / ou 'ill need to come into lab mornin)and eenin) hours to collect 2ies


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#nstructors

• Lecturer

– Diane 4"illane 5ds"ill( 678-7-19• !:ce hours Thursday 1081-;00 "m( 13,< =4B

•

GSIs and their Ofce hours held in 3131 USB – Din) >e( Wednesday 10 am81;00 "m

– 4usanna Cam"bell( Wednesday .8,;00 "m

– 4urabhi ?a$aram( Thursday .8,;00 "m

–

Chris 4i&uentes( @riday 11 am8-;00 "m


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Gradin) olicy

Grade Scale

• 360800 "ts % ran)e

• 3-083.E "ts B ran)e

• -,0831E "ts C ran)e• -08-7E "ts D ran)e

• F -3E "ts <

• #& necessary( these cutoHs 'ill be cured Nocurin) is done until the com"letion o& thecourse


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Grades/00 oints Total

• Iuies – 100 "ts( -.K o& )rade

• Drosophila ro$ect

– 16 "ts( 1K o& )rade• C. elegans ro$ect

– .0 "ts( 1-.K o& )rade

• >ome'or+ – 60 "ts( 1.K o& )rade

• Laboratory 'or+ – -6 "ts( 6.K o& )rade


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Iuies

• There are t'o Iuies )ien durin) thelast hour o& your laboratory "eriod – @ebruary --8-3rd

– %"ril 8.th

• #& you reuire s"ecial accommodations(you must brin) the a""ro"riate

documentation to Dr 4"illane at least -'ee+s "rior to the *rst ui


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Course Canas 4ite

• Lab Manual –


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4ub$ect Matter

• Drosophila "ro$ect – #denti&y "otential re)ulators o& a cell8cell

communication "ath'ay 5Win)less or Wnt si)nalin)9• #denti&y the chromosomal location o& random trans"oson

insertions that aHect the Win)less "ath'ay

• =se C? &ollo'ed by DN% seuencin) to more "reciselyidenti&y the trans"oson insertion 'ith the )oal o& *ndin)"otential )enes inoled in the Win)less "ath'ay

• Caenorhabditis elegans 5C. elegans9"ro$ect – Genetic ma""in) o& a mutation usin) molecular

mar+ers


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>o' are 'e )oin) to identi&y re)ulators o& W) si)nalin)

1 4tart 'ith a "henoty"e that results &rom oereA"ressionWin)less

- % 4creen 'as "er&ormed to identi&ytrans"oson insertions that su""ress the"henoty"e

!ereA"ressiono& W) in the

eye causes asmall


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The search &or re)ulators o& W) si)nalin) usestrans"oson

5G4OP9 insertions that allo' nearby )enes to be

actiated in an inducible manner4tretch o& 2y)enomic DN%

#nsert a construct5"G4OP9

G4OP

We hae many diHerent 2y stoc+s 'ith random G4OP insertionsMore on this later

The insert causes transcri"tional actiation o&

)enes oneither side o& the insert( )enes that may be

inoled in the W) si)nalin) "ath'ay


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#ntroduction to Drosophila

melanogaster


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Why =se Drosophila

• 4mall sie

• 4hort li&e cycle 51081 days9

•

>i)h re"roductie rate 5a sin)le&emale can lay 008.00 e))s in 10days9

•


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Droso"hila Li&e Cycle


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Becomin) @amiliar 'ithDroso"hila

horaA has three se)ments( each 'ith a "air o& le)s The 'iattached to T-( the haltere to T3


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%bdominal 4e)ments in Drosophila


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Droso"hila Chromosomes


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Droso"hila chromosomes

4econd "air

Third "air

4eA chromosome

@ourth "air

Di"loid-n Q ,


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Drosophila Cultures s Drosophila

Crosses

• Cultures – Males and &emales are o& the same

)enoty"es

–Ree"s a stoc+ o& 2ies &or &uture uses

• Crosses – Males and &emales are o& diHerent

)enoty"es

– =sed &or eA"erimental "ur"oses

– ou 'ill uses crosses to ans'er uestionsabout your un+no'n insertions


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Males s @emales( cont

Males hae sexcombs on the *rst

set o& le)s#m"ortant &or matin)


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The Best Way to 4eA @lies

• The male genitalclasers are themost accurate 'ay todistin)uish a male&rom &emale

• This is 'hat youshould do in thelab

• Could con*rm 'ithseA combs( but ta+estoo much time

ou tube ideo; htt";'''youtubecom'atchQ&Sla&h@$ec


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4ymbolism =sed• Wild Ty"e

–

We use the symbol U to denote 'ildty"e

– Can 'rite U ty"e

• Mutant

– %ny heritable deiation &rom 'ild ty"e ismutant

– We use an abbreiation o& the name(

such as ' &or 'hite 5'hite eye colormutation9

– Lo'er case letter &or a recessiemutation

– = er case letter &or a dominant


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Nomenclature continued

• Commas are used to se"arate traitson the same chromosome

• 4emi8colons are used to se"aratetraits on diHerent chromosomes

'' UU LyTM6B( 4b

What does this mean


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'' UU LyTM6B( 4b


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'' UU LyTM6B( 4b

The recessie 'hitemutation is located onthe V chromosome

This is a &emale


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The semi8colon se"arates the chromosomes

'' UU LyTM6B( 4b


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'' UU LyTM6B( 4b

Ly is a dominant mutationlocated on one co"y o& the3rd chromosome


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'' UU LyTM6B( 4b


TM6B is a dominant mutationlocated on the other co"y o&

The 3rd chromosome


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'' UU LyTM6B( 4b


TM6B is a dominant mutationlocated on the other co"y o&

The 3rd chromosome

4b is a dominant mutationlocated on the other co"y o

The 3rd chromosome( thesame chromosome as TM6


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Lab This Wee+

• Learn ho' to "ro"erly use "i"ettors

• Collect 30 2ies to be used &or GenomicDN% isolation

• Learn ho' to handle Droso"hila – =se the C0- system to anesthetie 2ies

• "er# imortant$$$

– #denti&y 2y body "arts and sta)es o& the li&ecycle

– Become com%ortable searating malesand %emales$


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Will # be 'or+in) alone

• @or the Droso"hila ro$ect – ou 'ill 'or+ in "airs to collect data

5some control data 'ill be shared 'ith

the class9 – >o'eer( all 'ritten re"orts and

home'or+ must be done indiidually

•

@or the C ele)ans ro$ect – ou 'ill 'or+ indiidually to collect data

and com"lete re"orts and home'or+

1 introductionn

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