TY - JOUR
T1 - [23] The Use of the T4 DNA Polymerase in Identification of 3′ Terminal Nucleotide Sequences of Duplex DNA
AU - Englund, P. T.
AU - Price, S. S.
AU - Weigel, P. H.
N1 - Funding Information:
This work was supported by the National Institutes of Health (Grants Nos. GM 16585-03 and CA 13602-04). P.T,E. received a Faculty Research Award from the American Cancer Society. S.S.P. and P.H.W. are supported by N.I.H. Training Grant No. 5T01 GM00184-13. Nancy Catterall and Patricia Clinkenbeard provided expert technical assistance.
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 1974/1/1
Y1 - 1974/1/1
N2 - The T4 DNA polymerase can be used to determine the sequence of several nucleotides at the 3' termini of duplex DNA molecules. The DNA is nonradioactive and the single deoxynucleoside triphosphate is labeled in the α-position with 32P. The T4 DNA polymerase attacks the 3' terminus of the DNA and sequentially releases mononucleotides. Aside from its utility in sequencing, the specific labeling of 3' termini of DNA by means of the T4 polymerase may prove to be as useful as has been the labeling of 5' termini by means of polynucleotide kinase. For example, it would be possible to exchange the two 3'-terminal nucleotides of a DNA molecule with labeled nucleotides. This reaction would introduce no net covalent changes into the molecule, yet each strand would contain a 3' terminal label. It is also possible, by using one triphosphate that is radioactive and one that is nonradioactive, to incorporate a label into one strand, but not into the other.
AB - The T4 DNA polymerase can be used to determine the sequence of several nucleotides at the 3' termini of duplex DNA molecules. The DNA is nonradioactive and the single deoxynucleoside triphosphate is labeled in the α-position with 32P. The T4 DNA polymerase attacks the 3' terminus of the DNA and sequentially releases mononucleotides. Aside from its utility in sequencing, the specific labeling of 3' termini of DNA by means of the T4 polymerase may prove to be as useful as has been the labeling of 5' termini by means of polynucleotide kinase. For example, it would be possible to exchange the two 3'-terminal nucleotides of a DNA molecule with labeled nucleotides. This reaction would introduce no net covalent changes into the molecule, yet each strand would contain a 3' terminal label. It is also possible, by using one triphosphate that is radioactive and one that is nonradioactive, to incorporate a label into one strand, but not into the other.
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U2 - 10.1016/0076-6879(74)29027-X
DO - 10.1016/0076-6879(74)29027-X
M3 - Article
C2 - 4855152
AN - SCOPUS:0016013516
VL - 29
SP - 273
EP - 281
JO - Methods in Enzymology
JF - Methods in Enzymology
SN - 0076-6879
IS - C
ER -