Plant DNA digests - Michigan State University Plant DNA digests Plant Plasmid CSS451 digests Plant...

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Plant DNA digests

Plasmid DNA digests Plasmid DNA digests

Plant DNA digests CSS451 Plant DNA digests

Plant Plasmid

CSS451 digests

Plant Plasmid

Southern AnalysisOptimizing DNA digest concentrations

2 4 8 12 16 20 24 ug

Estimating Yield of Dig-labeled Probe

1:5 1:50 1:500 1:5000 1:50,000

Plasmid DNA Southern Analysis

Bt-cry52244 bp

2.0kb1.9kb

21kb

5.1kb4.9kb

A

B

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

M NT LM L235 C3 80-1

Southern Analysis

Copynumber

Integration

NT LM L23 C3 80-11 2 4 5 6 7 8 9 10 11 12 13 14 15

4.0kb

3

Northern Analysis

Southern Analysis

Non-specificBinding of probebecause wrong buffer was used.

Non-bindingOf Probebecause of poorHybridization.

Image of Whole membrane

Dig-labeled size marker

Southern Analysis:Light-exposed film

Southern Analysis:Light-exposed film

cryV-BtPVYcp

RAPD Analysis of Bt-cry5/PVYcp transgenic potatoes

1 2 3 4 5 6 7 8

1.2kb

2.2kb A

B

1 2 3 4 5 6 7 8

Southern Analysis of Bt-cry5/PVYcp transgenic potatoes:Integration

1 2 3 4 5 6 7 8

3.5kb

4.2kb

4.9kb5.1kb

Southern Analysis of Bt-cry5/PVYcp transgenic potatoes:Copy Number

1.2kb

2.2kb A

B

1 2 3 4 5 6 7 8 9

1 2 3 4 5 6 7 8 9

Northern Analysis of Bt-cry5/PVYcp transgenic potatoes:Leaf tissue

1.2kb

2.2kb A

B

1 2 3 4 5 6 7 8

1 2 3 4 5 6 7 8

Northern Analysis of Bt-cry5/PVYcp transgenic potatoes:Tuber tissue

125kD

79.6kD

1 2 3 4 5 6 7 8 9 10 11 12 13

CRYV

Western Analysis of Bt-cry5 Potatoes

1 2 3 4 5 6 7 8 9 10 11 12 13 14 1516

500 bp

456 bp

256 bp

PVY

Cry1a

NPTII

Progeny analysis of crosses between PVYcp and Bt-transgenic potatoes:

PCR results

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 1621kb

21kb

5.1kb

5.1kb

4.2kb

4.2kb

3.5kb

3.5kb

PVY

Cry1a

Progeny analysis of crosses between PVYcp and Bt-transgenic potatoes:

Southern Analysis results

Fingerprint Analysis of Potato Varieties