Biochemistry
8th Edition
ISBN: 9781464126109
Author: Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr., Lubert Stryer
Publisher: W. H. Freeman
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Chapter 5, Problem 13P
Interpretation Introduction
Interpretation:
The information that is revealed by the given result about the structure of the encoded protein is to be stated.
Concept introduction:
DNA stands for deoxyribonucleic acid, is a biological macromolecule. DNA contains double helical strands along with the complementary base pairs. The four complementary bases of DNA are adenine (A), thymine (T), guanine (G) and cytosine (C).
The reaction that is used to make the duplicates of a particular DNA segment is known polymerase chain reaction (PCR).
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The short DNA shown below is to be sequenced. Using your knowledge of how the Sanger method
works, in the gel diagram, draw in the bands that will appear when DNA polymerase is added to the
reaction along with the four different nucleotide mixtures indicated. Note that some of these mixtures are
not what would normally be used in a sequencing reaction. Dideoxynucleatides (ddNTPs) are added in
relatively small amounts. The asterisk represents a radioactive label.
*5'
- 3'-ОН
3' –
-- ACGACGCAGGACATTAGAC-5'
Nucleotide mixtures:
A. DATP, DTTP, dCTP, DGTP, ddTTP (given)
B. DATP, ATTP, dCTP, AGTP, ddATE
C. dTTP, dGTP. ACTP, ddCTP, ddATP
D. DATP, dCTP, dTTP, ddGTP
A
в с D
| || ||
After restriction enzymes cut, they contain unpaired bases. Type II restriction enzymes leave ends that may be 5' overhanging, 3' overhanging, or blunt. In all cases each end is left with a 3' OH and a 5' phosphate. All blunt ends, and any complementary overhanging ends may be re-ligated with T4 DNA ligase, as long as at least one 5'- phosphate is present.
In the tables below G^AATTC means that the end after cutting with enzyme will be:
-----G 3'
-----CTTAA 5'
GTGCA^C means that the end will be:
-----GTGCA 3'
-----C 5'
Which RE’s from table below have a 5’ overhang? Which ones have a 3’ Overhang?
AccI
GT^CGAC
BamHI
G^GATCC
ClaI
AT^CGAT
NsiI
ATGCA^T
PstI
CTGCA^G
BglII
A^GATCT
TaqI
T^CGA
PCR primers
Below is a 300 base pair fragment of DNA. The top strand is written in the 5' to 3' direction. The bottom
strand is written 3' to 5'. There are also two primer sequences; both primers are written 5' to 3'. Note
that we are displaying a double-stranded DNA fragment, but primers will only bind to one of the two
displayed strands.
5' ACCGȚAGCTATATGCTATCGTGACGTATCGGCGCATTAAȚCGGGATCGAT 3
50
3'
TGGCÁTCGATATACOATAGCACTOCATAGCCGCGTAATTÀGCCCTAGCTÀ 5'
5' AGCTÇGCTAGCAGGAGAGAȚATCGÇTCATAGCTCCGATCGATGCCGCTAA 3
3' TCGAGCG ATCGTCCTCTCTÁTAGCGAGTATCGAGÓCTAGCTACGGCGATİ 5'
100
5' TATAGCTCTÇTGCGGATATÇGCATATACCẠ AGGCCCTACGTATGTAGCTA 3
150
3' ATATČGAGAGACOCCTATAGCGTATATGGTTCCGGGATGČATACATCGAŤ 5'
5 TGCGTATATÇGGAGAGTCCTGGATATGGAGCTTGACTGCAGAGAGCTCGA 3
200
3' ACGCÁTATAGCCTCICAGGÁCCTATACCTCGAACTGACGTCTCTCGAGCT 5'
5' TATGCGCTTAGGCCGTATATGCTTGGGGAAAGCTCTATGTATGCTATGTG 3
3. ATACGCGAATCCGGCATATACGAACCCCTÍTCGAGATACATACGATACAC 5'
250
5' TGCATGTGCTATGCAACGTTCOGATTGCGȚAGCAGTAATAGCGCCGATTG 3
300
3'…
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