Bio97_09+L20

Bio97_09+L20 - Human gene mapping OFFI C HOURS TODAY 12-3 E...

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1 Human gene mapping OFFICE HOURS, TODAY 12-3 McGaugh Hall 4213
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2 1. Pedigree 2. collect DNA from all individuals in pedigree 3. Restriction digest 4. Gel electrophoresis 5. Southern Blot 6. Probe for change in restriction site at known genomic location 6. Probe for change in length of an SSR at known genomic location 3. PCR amplify an SSR at known genomic location 4. Gel electrophoresis * * * * * * * *
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©2003 C. Brachmann 3 SSR10 SSR190 SSR8
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©2003 C. Brachmann 4 SSR89 SSR89 5’ gggcgg gcgttctcgcatagt ccc -varying TG-- aaacctgtgacccttgaatg 3’ 3’ cccgcccgcaagagcgtatcaggg -varying AC-- tttgg acactgggaacttac 5’ 5’gggcgggcgttctcgcatagtccc tg-- 88 -- aaacctgtgacccttgaatg 5’gggcgggcgttctcgcatagtccc tg-- 69 -- aaacctgtgacccttgaatg 5’gggcgggcgttctcgcatagtccc tg-- 61 -- aaacctgtgacccttgaatg 5’gggcgggcgttctcgcatagtccc tg-- 49 -- aaacctgtgacccttgaatg 5’gggcgggcgttctcgcatagtccc tg-- 42 -- aaacctgtgacccttgaatg 5’gggcgggcgttctcgcatagtccc tg-- 38 -- aaacctgtgacccttgaatg allele 1 allele 2 allele 3 allele 4 allele 5 allele 6 pcr primer sequences in bold
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©2003 C. Brachmann 5 ©2003 C. Brachmann Once you’ve identified an SSR that shows linkage, you know the genomic region for phd Then sequence genes in that region. Expect that phd people have a mutation relative to normals in the phd gene. Can use the information of phd linkage with allele 6 of SSR89 to identify others in the population likely to have phd
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©2003 C. Brachmann 6 Applications of human genetic mapping: Clone genes implicated in human disease: Family pedigree analysis important. Criminal investigations: With >>5000 polymorphisms known in humans, RFLPs and SSRs are equivalent to genetic “finger prints”. Paternity tests: Inheritance patterns of RFLPs can resolve paternity, identify corpses or solve other “missing” cases 23andME:550,000 SNPs linked to traits/disease
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©2009 C. Brachmann 7 ©2003 C. Brachmann Paternity testing Set 1 and 2 contain results from two different paternity cases. In each case, the lanes contain DNA fragments from the following sources: M = Mother C = Child A = Alleged father Q: Is the alleged father (A) the true biological father?
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©2003 C. Brachmann 8 DNA Fingerprinting In this example, an SSR was detected by restriction enzyme
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Bio97_09+L20 - Human gene mapping OFFI C HOURS TODAY 12-3 E...

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