F13 lecture 6 for post

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Unformatted text preview: •  •  •  5 individuals (assume haploids or homozygous diploids) EcoR1 HindIII 1 EcoR1 1 1 kb EcoR1 HindIII 3 kb EcoR1 2 EcoR1 4kb -3kb -- EcoR1 3 EcoR1 HindIII EcoR1 1kb -- 4 EcoR1 5 EcoR1 isolate genomic DNA from individuals PCR amplify fragment of interest containing variable restric1on site digest with variable restric1on enzyme only run on gel EcoR1 HindIII EcoR1 2 3 4 5 RFLPs are co-dominant markers =can tell all three genotypes RAPD markers •  Random Amplifica1on of Polymorphic DNA –  PCR with 1 primer of arbitrary sequence –  a series of fragments are produced, most invariable, some variable between individuals •  these are inherited like alleles, either band, or no band –  Have problems with reproducability – no longer used much http://www.informatics.jax.org/silver/figures/figure8-9.shtml RAPD markers are are dominant markers two alleles: band produced no band produced =heterozygote for the band looks the same as homozygote (although quantification is possible in some cases) AFLP markers •  Amplified Fragment Length Polymorphism ①  isolate genomic DNA and digest with restric1on enzyme ②  ligate (a)ach) “adaptors” onto “s1cky ends” of restric1on- digested DNA fragments ③  perform PCR with primers complementary to adaptor sequences plus 1 or a few nucleo1des into the restric1on fragments •  results in PCR amplifica1on of a subset of the fragments in the sample ④  run products on gel – many bands are variable between individuals – some are inherited as band or no band markers...
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This document was uploaded on 02/27/2014.

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