After transduction we move to mutagenesis analysis. Using the transduction plate,PCR I found that lacZ,fliE,nagC,ompA all show successful transduction and successful PCR. But for DNA sequencing some clonies are not correct. Expected size of PCR are lacZ 304,fliE 255,rpoA 420,nagC 329,ompA 731,lexA 350,kan cassette 549,tet cassette 265. Please indicate which mutation were successful and success why some were not from the size about. indicate at what stage any of the mutagenesis failed.

Human Anatomy & Physiology (11th Edition)
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Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
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After transduction we move to mutagenesis analysis. Using the transduction plate,PCR I found that lacZ,fliE,nagC,ompA all show successful transduction and successful PCR. But for DNA sequencing some clonies are not correct. Expected size of PCR are lacZ 304,fliE 255,rpoA 420,nagC 329,ompA 731,lexA 350,kan cassette 549,tet cassette 265. Please indicate which mutation were successful and success why some were not from the size about. indicate at what stage any of the mutagenesis failed.
Batch Gene Colony
number
1
2
3
number
1
23
4
ompA
Lacz
fliE
naqC
4
1
1
2
3
1
234
2
Correct / incorrect
correct
incorrect
incorrect
correct
incorrect
incorrect
incorrect
correct
correct
correct
correct
correct
Evidence (notes)
Transcribed Image Text:Batch Gene Colony number 1 2 3 number 1 23 4 ompA Lacz fliE naqC 4 1 1 2 3 1 234 2 Correct / incorrect correct incorrect incorrect correct incorrect incorrect incorrect correct correct correct correct correct Evidence (notes)
nagC primers
M L1 L2 L3 L4
|||||||||||||
lacZ primers
M L1 L2 L3 L4
||||||||||||
fliE primers
M L1 L2 L3 L4
ompA primers
M L1 L2 L3 L4
tet primers
M L1 L2 L3 L4
||||||||||||
kan primers
M L1 L2 L3 L4
||||||||||||||
kan primers
M L1 L2 L3 L4
kan primers
M L1 L2 L3 L4
| | || | ||| | |||| 3
bp
10,000
-8,000
-6,000
-5,000
-4,000
-3,000
-2,500
-2,000
-1,500
¹-1,000
-750
-500
-250, 253
1.2% agarose
k
I=
L
Transcribed Image Text:nagC primers M L1 L2 L3 L4 ||||||||||||| lacZ primers M L1 L2 L3 L4 |||||||||||| fliE primers M L1 L2 L3 L4 ompA primers M L1 L2 L3 L4 tet primers M L1 L2 L3 L4 |||||||||||| kan primers M L1 L2 L3 L4 |||||||||||||| kan primers M L1 L2 L3 L4 kan primers M L1 L2 L3 L4 | | || | ||| | |||| 3 bp 10,000 -8,000 -6,000 -5,000 -4,000 -3,000 -2,500 -2,000 -1,500 ¹-1,000 -750 -500 -250, 253 1.2% agarose k I= L
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