Dna Sequencing by Sanger Method
Dna Sequencing by Sanger Method
Dna Sequencing by Sanger Method
to
Bioinformatics
BIN101
Tube A
15% ATP
25% GTP
25% TTP
25% CTP
10% ddATP
Sanger Sequencing Setup
AAT C T G G G C TA C T C G G G C G T T
C G CA
Tube A
15% ATP
25% GTP
25% TTP
25% CTP
10% ddATP
Sanger
Sequencing
AAT C T G G G C TA C T C G G G C G T T
C G CA
Setup A G C C C G CA
Tube A
15% ATP
25% GTP
25% TTP
25% CTP
10% ddATP
Sanger
Sequencing
AAT C T G G G C TA C T C G G G C G T T
C G CA
Setup A G C C C G CA
AT G A G C C C G CA
Tube A
15% ATP
25% GTP
25% TTP
25% CTP
10% ddATP
Sanger
Sequencing
AAT C T G G G C TA C T C G G G C G T T
C G CA
Setup A G C C C G CA
AT G A G C C C G CA
Tube A
15% ATP
25% GTP
AC C C GATGAG C C C
GGCC
GAT
25% TTP
25% CTP
10% ddATP
Sanger
Sequencing
AAT C T G G G C TA C T C G G G C G T T
C G CA
Setup A G C C C G CA
AT G A G C C C G CA
Tube A
15% ATP
25% GTP
AC C C GATGAG C C C
GGCC
GAT
25% TTP
A G A C C C G A T G A G C C C G CA 25% CTP
10% ddATP
Problem:
1. The following DNA fragment was sequenced by the Sanger method. The red asterisk
indicates a fluorescent label.
A sample of the DNA was reacted with DNA polymerase and each of the nucleotide
mixtures (in an appropriate buffer) listed below. Dideoxynucleotides (ddNTPs) were added
in relatively small amounts.
1. dATP, dTTP, dCTP, dGTP, ddTTP
2. dATP, dTTP, dCTP, dGTP, ddGTP
3. dATP, dCTP, dGTP, ddTTP
4. dATP, dTTP, dCTP, dGTP
The resulting DNA was separated by electrophoresis on an agarose gel, and the fluorescent
bands on the gel were located. The band pattern resulting from nucleotide mixture
1 is shown below.
Assuming that all mixtures were
run on the same gel,
what did the remaining lanes
of the gel look like?