Using the data. Which genes have successful transduction?successful PCR?

Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
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Chapter1: The Human Body: An Orientation
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Using the data. Which genes have successful transduction?successful PCR?successful DNA sequencing?
Table 4. Transduction Plates Result
Number of colonies
74
0¹
Plate
E. coli-only
nagC phage only
rpoA phage only
nagC+ E. coli repeat 1
nagC+ E. coli repeat 24
nagC+E. coli repeat 24
rpoA + E. coli repeat 1
rooA + E. coli repeat 24
rpoA + E. coli repeat 34¹
Buffer only
Gene
Size of
gene (bp)
lacz
3075
fliE
315
rpoA
990
nagC
1221
ompA
1041
lexA
609
kan cassette 816
Forward Primer
5' to 3'
GTTAACCGTCACGAGCATC
ACAGGGGATTGAAGGGGTTA
CGTTCAGGAAGATATCC
CGCTGCCAACGCTGC
0
12
104
144
GGCCCGACCCATGAAAAC
GGCACGCAAAGGCGTTATTGA
0¹
04
0
04
Task 2: Find where the primers bind to in the gene sequences on Moodle (kan has been
provided for you as a reference)
e
Reverso primer
5' to 3
P
2
2
e
←
2
P
P
32
43
TGATCACACTCGGGTGATTAC
CGCACCTGAATCCCCATTTGC
GGATCGATTGTGCCGTTGGTTT
||
Expected size of
PCR product (bp)
325
250
GCTCAGAACGTACCACCG
GCCGGGATACCTTTGG
GTTCGACTTTATTGCCCTG
CCTGGTATCGGTCTGC
GGAAGCCCGATGCGCCAGAG
tet casette
1148
CTATAAACCGCACTGTGA
GTCCTGGATTCCTCAGGT
The reverse primer is the reverse compliment-remember to reverse compliment when looking for it in the sequences. Kan has been
completed for you
375
549
Transcribed Image Text:Table 4. Transduction Plates Result Number of colonies 74 0¹ Plate E. coli-only nagC phage only rpoA phage only nagC+ E. coli repeat 1 nagC+ E. coli repeat 24 nagC+E. coli repeat 24 rpoA + E. coli repeat 1 rooA + E. coli repeat 24 rpoA + E. coli repeat 34¹ Buffer only Gene Size of gene (bp) lacz 3075 fliE 315 rpoA 990 nagC 1221 ompA 1041 lexA 609 kan cassette 816 Forward Primer 5' to 3' GTTAACCGTCACGAGCATC ACAGGGGATTGAAGGGGTTA CGTTCAGGAAGATATCC CGCTGCCAACGCTGC 0 12 104 144 GGCCCGACCCATGAAAAC GGCACGCAAAGGCGTTATTGA 0¹ 04 0 04 Task 2: Find where the primers bind to in the gene sequences on Moodle (kan has been provided for you as a reference) e Reverso primer 5' to 3 P 2 2 e ← 2 P P 32 43 TGATCACACTCGGGTGATTAC CGCACCTGAATCCCCATTTGC GGATCGATTGTGCCGTTGGTTT || Expected size of PCR product (bp) 325 250 GCTCAGAACGTACCACCG GCCGGGATACCTTTGG GTTCGACTTTATTGCCCTG CCTGGTATCGGTCTGC GGAAGCCCGATGCGCCAGAG tet casette 1148 CTATAAACCGCACTGTGA GTCCTGGATTCCTCAGGT The reverse primer is the reverse compliment-remember to reverse compliment when looking for it in the sequences. Kan has been completed for you 375 549
nagC primers
M L1 L2 L3 L4
|||||||||||||
lacZ primers
M L1 L2 L3 L4
fliE primers
M L1 L2 L3 L4
I
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
bp
-10,000
-8,000
-6,000
-5,000
-4,000
-3,000
-2,500
-2,000
-1,500
-1,000
-750
-500
-250, 253
1.2% agarose
k
I=
Transcribed Image Text:nagC primers M L1 L2 L3 L4 ||||||||||||| lacZ primers M L1 L2 L3 L4 fliE primers M L1 L2 L3 L4 I 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 bp -10,000 -8,000 -6,000 -5,000 -4,000 -3,000 -2,500 -2,000 -1,500 -1,000 -750 -500 -250, 253 1.2% agarose k I=
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