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Molecular_Technologies_2_Week_9_Fall_2009

Molecular_Technologies_2_Week_9_Fall_2009 - Week9Notes...

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Week 9 Notes Animal Breeding  Fall 2009
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Extraction of DNA can come from almost  any tissue or body fluid Blood Muscle Saliva Semen Skin Concentration of DNA will vary from tissue  and extraction procedure
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Importance of DNA extraction DNA repositories Research populations Identification of carriers of alleles through DNA and  pedigree analysis Collaborative efforts of multiple researchers
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DNA from a “foundation” population and  every individual born in the population in  subsequent generations, or brought into  the population from the outside Multi-generational analyses of inheritance  Multi-generational analyses of successful  selection Large population of animals to conduct marker  association studies
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Genetic marker  A detectable gene or DNA fragment used to  identify alleles
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Microsatellites DNA which is composed of many tandem  repeats of short basic repeating units Typically in non-coding regions RFLP Single Nucleotide Polymorphisms SNP’s Single base pair change in DNA sequence
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GAACT A GCTTG GAACT G GCTTG SNP
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CCGTAT CACACA GTCAAC CCGTAT CACACACA CAGTCAAC Microsatellite
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Taking a small amount of extracted DNA  and creating a larger concentration for  research purposes Mimics DNA replication in a laboratory setting Forward and Reverse Primers dNTP’s (nucleotides) Buffer Mg Taq Polymerase Assembles new DNA 
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Denaturing Step Heating and unwinding (melting) of DNA Annealing Step Reaction is cooled Primers anneal to DNA DNA synthesis begins Extension/elongation New DNA synthesis via Taq begins and extension of the  primer sequence  Final elongation Safety step to ensure all single stranded DNA is elongated Final hold Allows for short term storage of reaction
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DNA    C/C    T/T  T/T    T/T      T/T   T/T     C/C      T/T  T/T   T/T    C/T     C/C    T/T  T/T     T/T    T/T   T/T Ladder
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Utilizing up to 4 primer pairs to generate  DNA fragments Can generate genotypes for four “spots” of  interest  in a single reaction
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