2 SNPs Single nucleotide polymorphisms regions of DNA where one base pair is different. Occur evenly spread over all the DNA. 1/ 1000-3000 bp Detected by sequencing. If SNP occurs in a restriction enzyme site it generates an RFLP. Could be in coding or non-coding regions. Over 300000 human SNPs known and are being mapped. 3 SNP 4 RFLPs
Restriction fragment length polymorphism.
Any difference in the DNA that results in a restriction enzyme producing a different sized piece.
Mutation at a restriction site prevents recognition cutting.
Results in one band of larger DNA instead of 2 smaller ones.
Different numbers of repeats between restriction recognition sites also generates an RFLP
5 RFLPs www.ncbi.nlm.nih.gov/.../doc/TechRFLP.shtml DNA marker may be associated with genetic condition. 6 VNTRs and STRPs as RFLPs Minisatellites and Microsatellites
These are RFLPs because they are defined by or visible following restriction enzyme cuts.
Variable Number Tandem Repeats
Groups (10-100) of nucleotides repeated 2 100 times (depending on individual and locus).
Restriction sites on both sides of repeated DNA
The more repeats the longer the fragment.
Simple Tandem Repeat Polymorphisms
Shorter 2-9 nucleotides repeated
Small enough number for PCR amplification
Also called STRs SSLPs etc.
7 Use of VNTRs Restriction sites are on either side fragment length depends on number of repeats in between sites. 8 STRPs Primers for both sides of repeated region allow PCR amplification of DNA generates PCR products that differ in length depending on number of repeats. Becoming the standard method for DNA testing in forensics labs. Cheaper easier more sensitive. 9 Hardy-Weinberg meets Gil Grissom
Simple tandem repeats
13 have been chosen for use in forensic work
The 13 independently assort meaning they are on different chromosomes or far apart on the same.
Product law can be used
Each of the 13 have a number of different alleles
Alleles differ by number of repeats
Repeats vary from 3 to 5. vWA is a tetranucleotide.
Allele frequencies p1 p2 etc. for each allele
Humans are diploid have 2 alleles for each locus
10 STRs in forensics
Alleles in different ethnic and racial groups examined used as database. Panel of 13 different STRs are used. Because the odds of a particular combination of the 13 is product of the frequencies numbers like 1 in 10 billion can be generated. Hardy-Weinberg 2pq Band frequency 11 THE 13 CODIS STRs and probabilities of Identity http//expertpages.com/news/dna.htm 12 Allele frequencies for D1S80 among US population groups Chance of a white person being heterozygous for alleles 19 and 20 2 x 0.003 x 0.018 (One in 9259) 13 RAPD using PCR to find polymorphisms
Random amplified polymorphic DNA
Screen DNA from individuals by doing PCR with random short primers. (about 8-12 nucleotides)
By random chance primers will amplify many different sections of DNA.
Look for bands on gel that are not present in each individual tested.
avery.rutgers.edu/.../ archives/onions/rapd.html 14 RAPD using PCR to find polymorphisms-2
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