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Genetics & Genotyping

Genetics & Genotyping. By Kristin Gill & Daisy Robinton HC70AL Spring 2009. What are the possible genotypes?. Homozygous Wildtype Hemizygous Wildtype/T-DNA Homozygous T-DNA. What is the expected ratio of genotypes?. 1 : 2 : 1. Homozygous Wildtype. Homozygous T-DNA. Hemizygous.

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Genetics & Genotyping

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  1. Genetics & Genotyping By Kristin Gill & Daisy Robinton HC70AL Spring 2009

  2. What are the possible genotypes? • Homozygous Wildtype • Hemizygous Wildtype/T-DNA • Homozygous T-DNA

  3. What is the expected ratio of genotypes? 1 : 2 : 1 Homozygous Wildtype Homozygous T-DNA Hemizygous

  4. What are the primer interactions? Wildtype Product = FORWARD + REVERSE T-DNA Product = LBb1 + FORWARD OR LBb1 + REVERSE

  5. Structure of a Gene + Primers 5’ 3’ T T A T RV A A T A 5’ 3’ 3’ 5’ A C T G FW 3’ T G A C 5’ Given Primers from STOCKS Forward: 5’ ACTG 3’ Reverse: 5’ ATAA 3’ In Sequence Look For Forward: 5’ ACTG 3’ Reverse: 5’ TTAT 3’ Reverse Complement

  6. LBb1 What is a Knock-Out? 5 kb T-DNA Insert 5’ 3’ T T A T RV A A T A 5’ 3’ 3’ 5’ A C T G FW 3’ T G A C 5’ T-DNA genotype based on FORWARD Primer & LBb1 Primer Interaction

  7. LBb1 What is a Knock-Out? 5’ 3’ T T A T RV A A T A 5’ 3’ 3’ 5’ A C T G FW 3’ T G A C 5’ 5 kb T-DNA Insert T-DNA genotype based on REVERSEPrimer & LBb1 Primer Interaction

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