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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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Presentation Transcript
genetics genotyping
Genetics & Genotyping

By Kristin Gill & Daisy Robinton

HC70AL Spring 2009

what are the possible genotypes
What are the possible genotypes?
  • Homozygous Wildtype
  • Hemizygous Wildtype/T-DNA
  • Homozygous T-DNA
what is the expected ratio of genotypes
What is the expected ratio of genotypes?

1 : 2 : 1

Homozygous Wildtype

Homozygous T-DNA

Hemizygous

what are the primer interactions
What are the primer interactions?

Wildtype Product = FORWARD + REVERSE

T-DNA Product = LBb1 + FORWARD

OR

LBb1 + REVERSE

structure of a gene primers
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

what is a knock out

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

what is a knock out1

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