MCDB101Bslideset8

MCDB101Bslideset8 - 1 The Direct Detection of Genotype...

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Unformatted text preview: 1 The Direct Detection of Genotype Distinguishes Individual Genomes (Chpt. 11) Four classes of DNA variation for direct detection of genotype Protocols used to determination of allelic DNA polymorphisms. Positional cloning Linkage mapping LOD scores Candidate gene analysis Confirmation of gene identification transgenics 283 Co In E Four classes of DNA polymorphisms 284 Co In E 2 Microsatellites 1 every 30,000 bp Repeated units 2 5 bp in length Mutate by replication error Mutation rate of 10-3 per locus per gamete Useful as highly polymorphic DNA markers Fig. 11.3 285 Co In E Minisatellites Repeating units 20- 100 bp long Total length of 0.5 20 kb 1 per 100,000 bp, or about 30,000 in whole genome Fig. 11.4 286 Co In E 3 Figure 11.5 Small Insertions and Deletions 287 Co In E DNA Polymorphism Detection Systems Single nucleotide polymorphic allele differences detected by RFLP, ASO, or single nucleotide primer extension. Southern Blot or PCR to detect size differences between alleles for mini- and microsatellites polymorphisms 288 Co In E 4 SNP detection using southern blots Restriction fragment length polymorphisms (RFLPs) are size changes in fragments due to the loss or gain of a restriction site Fig. 11.6 289 Co In E SNP detection by PCR Must have sequence on either side of polymorphism Amplify fragment Expose to restriction enzyme Gel electrophoresis e.g., sickle-cell genotyping with a PCR based protocol Fig. 11.7 290 Co In E 5 SNP detection by ASO Very short probes (<21 bp) specific which hybridize to one allele or other Such probes are allele-specific oligonucleotides (ASOs)18 Fig. 11.8 291 Fig. 11.8 Co In E ASOs can determine genotype at any SNP locus Fig. 11.9 a-c 292 Co In E 6...
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MCDB101Bslideset8 - 1 The Direct Detection of Genotype...

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