dna fp...mahtab

7/30/2019 Dna Fp...Mahtab

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MD.MAHTABM.Sc.

BIOTECHNOLOGY


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Era of DNA Technology:Information through

DNA

The molecule is so beautiful. Its glory was reflected on Francis and me.

- James Watson on DNA


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Biometricsuniquely recognizing humans


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The blueprints (genetic information) formaking proteins is stored within our DNA. It is

the job of DNA to control the order in whichthese 20 different amino acids are put together.

The DNA of every human being on the planet is99.9% the same. It is the 0.1% that makes all

the difference!

Any type of organism can be identified byexamination of DNA sequences which is

unique to that species.

The molecule of life: DNA


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Technology Transition

DNA Fingerprinting Dr. Alec Jeffrey 1985

DNA Profiling FBI (RFLP) 1988

PCR STRs 1993

Mitochondrial DNA - 1996

SNPs

Chips


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YOUR DNA: Your Ultimate Genetic Bar Code

DNA Fingerprinting is a methodwhere:

a persons genetic traits, genes, are

used to make specific strings of DNA

letters that are cut into patterns ofshorter strings separated by length

these banding patterns canidentify a unique human being!

YOUR DNABandingPattern Will Identify

YOU! Image of a DNA fingerprint


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Restriction Fragment Length

Polymorphism (RFLP)

Detects a single base pair change in DNA

Must occur within a restriction enzyme cleavage sequence to be

visible

It is the length differences associated with DNA strandsor RFLPs that allow one to distinguish one person fromanother

Often used in disease screening such as in the detection ofsickle cell anemia


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RFLP


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What creates this unique pattern?

Satellite DNA: repetitive DNA sequence.

Macrosatellite: core sequence 100 to 6500bp

Minisatellite: core sequence of 10-20bp repeatedmultiple times

Microsatellite: small arrays of tandem repeats of2 to 4bp in length


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Repeats of Satellite DNA

Repeat units vary in length from 2bp to long stretches of6000bp or more

These repeat units are lined up head to tail and composesatellite DNA and are interspersed throughout the genome

The number of units varies person to person

Thus these sequences are calledVNTRs (variable numberof tandem repeats)

A VNTR is a locus that is hyper variable due to a largenumber of alleles each characterized by a differentnumber of repeat units


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Southern blotting can be used to visualize the variation

Probes specific to the repeat unit are hybridized to DNAcut with a restriction enzyme that cuts just outside theVNTR

This allows for the difference in VNTR length to be

detected

Two commonly used probes are known as:

33.6 (AGGGCTGGAGG)18

31.5 (AGAGGTGGGCAGGTGG)29

These are multi-locus minisatellite probesand show about17 different DNA bands for each individual


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PCR amplification of VNTR

PCR is particularly useful in forensic analysis as it allowsminute amounts of DNA to be analyzed

DNA can be obtained from blood stains, semen, saliva, orhair roots

Instead of digesting the DNA PCR is used to amplify theVNTRs and the products are run on a gel and visualized by

stainingThis process requires primers that anneal just outside theVNTR


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Short Tandem Repeats (STR)

Are a variation on VNTRs, but use the smallest repeatsunits often only 2 to 4 bp in length

aatttttgtattttttttagagacggggtttcaccatgttggtcaggctgactatgga

gt

tattttaaggttaatatatataaagggtatgatagaacacttgtcatagtttagaacg

aa

ctaacgatagatagatagatagatagatagatagatagatagatagatagatagacag

at

tgatagtttttttttatctcactaaatagtctatagtaaacatttaattaccaatatt

tg

13 core loci of tetrameric repeats are tested together tomake a DNA profile

The sequence above is locus D7S280 which is located on

chromosome 7


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CriminalIdentification &Forensics

DNA fingerprints can be usedas biological evidence

Strands of DNA can be foundon hair, blood or semen.

DNA isolated from thoseevidence can be comparedthrough VNTR patterns.

Useful in solving crimes likemurder and rape.

Example: The scandal ofPresident Clinton with MonicaLewinsky
http://rds.yahoo.com/S=96062857/K=Lewinsky+and+Clinton/v=2/TID=I001_0/SID=w/l=II/SS=i/OID=ed8d134fe00a0c5a/R=1/*-http://images.search.yahoo.com/search/images/view?back=http%3a//images.search.yahoo.com/search/images%3fsrch=1%26p=Lewinsky%2band%2bClinton%26ei=UTF-8%26n=20%26fl=0&h=121&w=150&imgcurl=www.washingtonpost.com/wp-srv/images/homepg2/clinton_lewinsky_montage.jpg&imgurl=www.washingtonpost.com/wp-srv/images/homepg2/clinton_lewinsky_montage.jpg&name=%3cb%3eclinton%3c/b%3e_%3cb%3elewinsky%3c/b%3e_montage.jpg&p=Lewinsky+and+Clinton&rurl=http%3a//www.washingtonpost.com/wp-srv/politics/special/clinton/faq.htm&rcurl=http%3a//www.washingtonpost.com/wp-srv/politics/special/clinton/faq.htm&type=jpeg&no=1&tt=137http://rds.yahoo.com/S=96062857/K=DNA+fingerprinting/v=2/TID=I001_0/SID=w/l=II/SS=i/OID=cd44fa1707c4864a/R=3/*-http://images.search.yahoo.com/search/images/view?back=http%3a//images.search.yahoo.com/search/images%3fp=DNA%2bfingerprinting%26ei=UTF-8%26cop=mss%26tab=3%26b=1&h=208&w=106&imgcurl=www.vetark.co.uk/Images/DNA%2520fingerprint.jpeg&imgurl=www.vetark.co.uk/Images/DNA%2520fingerprint.jpeg&name=%3cb%3eDNA%3c/b%3e+fingerprint.jpeg&p=DNA+fingerprinting&rurl=http%3a//www.vetark.co.uk/birdDNApages.html&rcurl=http%3a//www.vetark.co.uk/birdDNApages.html&type=jpeg&no=3&tt=364http://en.wikipedia.org/wiki/File:Codis.svg


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CODISCombined DNA Index

System

National software developed by the FBI

Distributed to local, state, and national crime labs

All 50 states mandate inclusion of DNA fingerprint (if available)from violent and sexually motivated crimes

Mostly a database of STR regions

Thousands of matches have led to the capture of criminals thatotherwise would not have been caught

This has led numerous people to suggest a national DNA database

that would include only polymorphism information
http://en.wikipedia.org/wiki/File:Codis.svg


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Parentage tests

determine if the alleged father of a child is

the biological father

The child (C) will share one band with thebiological mother (M) and one band with

alleged father #1 (AF1), the biological father.

No bands are shared between the child andalleged father #2 (AF2), the excluded male.


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References DNA Fingerprinting Using Amplified Fragment Length Polymorphisms (RFLP)

By: Heidi Chial, Ph.D. (Write Science Right) 2008 Nature Education

Citation: Chial, H. (2008) DNA fingerprinting using amplified fragment length polymorphisms (AFLP): No genomesequence required. Nature Education1(1)

Forensics, DNA Fingerprinting, and CODIS

By: Karen Norrgard, Ph.D. (Write Science Right) 2008 Nature Education

Citation: Norrgard, K. (2008) Forensics, DNA fingerprinting, and CODIS. Nature Education1(1)

"CODIS National DNA Index System". Fbi.gov. Retrieved 2010-04-03.

Codis Statistics, 06/2008,

(http://www.fbi.gov/hq/lab/codis/clickmap.htm)

Kijkmagazine, 01 January 2009

Use of DNA in Identification". Accessexcellence.org. Retrieved 2010-04-03.

"Restrictions on use and destruction of fingerprints and samples". Wikicrimeline.co.uk. 2009-09-01. Retrieved 2010-04-03.

Lewinsky scandal", The Columbia Encyclopedia, Sixth Edition, 2008,As Retrieved 2010-02-09

John M. Butler, Forensic DNA Typing: Biology, Technology, and Genetics of STR Markers, Second Edition, Academic

Press, 2005.

DNA Fingerprint Analysis of Three Short Tandem Repeat (STR)

Loci for Biochemistry and Forensic Science Laboratory CoursesS

Received for publication, November 21, 2005, and in revised form, April 3, 2006 Kathleen McNamara-Schroeder,

Cheryl Olonan, Simon Chu, Maria C. Montoya, Mahta Alviri,

Shannon Ginty, and John J. Love

From the Department of Chemistry and Biochemistry, San Diego State University,

San Diego, California 92182-1030
http://www.fbi.gov/hq/lab/codis/national.htmhttp://www.fbi.gov/hq/lab/codis/national.htmhttp://www.fbi.gov/hq/lab/codis/national.htmhttp://www.accessexcellence.org/RC/AB/BA/Use_of_DNA_Identification.phphttp://www.wikicrimeline.co.uk/index.php?title=Identification_by_body_samples_and_impressionshttp://www.encyclopedia.com/topic/Lewinsky_scandal.aspxhttp://www.encyclopedia.com/topic/Lewinsky_scandal.aspxhttp://www.encyclopedia.com/topic/Lewinsky_scandal.aspxhttp://www.encyclopedia.com/topic/Lewinsky_scandal.aspxhttp://www.encyclopedia.com/topic/Lewinsky_scandal.aspxhttp://www.encyclopedia.com/topic/Lewinsky_scandal.aspxhttp://www.wikicrimeline.co.uk/index.php?title=Identification_by_body_samples_and_impressionshttp://www.accessexcellence.org/RC/AB/BA/Use_of_DNA_Identification.phphttp://www.fbi.gov/hq/lab/codis/national.htmhttp://www.fbi.gov/hq/lab/codis/national.htmhttp://www.fbi.gov/hq/lab/codis/national.htmhttp://www.fbi.gov/hq/lab/codis/national.htmhttp://www.fbi.gov/hq/lab/codis/national.htm


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