Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2',3'-dideoxyribonucleotide
About: This article is published in Biochemistry.The article was published on 1969-12-01. It has received 145 citations till now.
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TL;DR: A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.
Abstract: A new method for determining nucleotide sequences in DNA is described. It is similar to the “plus and minus” method [Sanger, F. & Coulson, A. R. (1975) J. Mol. Biol. 94, 441-448] but makes use of the 2′,3′-dideoxy and arabinonucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase. The technique has been applied to the DNA of bacteriophage ϕX174 and is more rapid and more accurate than either the plus or the minus method.
62,728 citations
TL;DR: Exonuclease function of DNA polymerase from Escherichia colis, discussing hydrolysis of polydeoxyribonucleotides and resistancy of oligonucleotide.
705 citations
TL;DR: If ASPCR is able to discriminate all allelic variation (both transition and transversion mutations), this method has the potential to be a powerful approach for genetic disease diagnosis, carrier screening, HLA typing, human gene mapping, forensics, and paternity testing.
Abstract: A rapid nonradioactive approach to the diagnosis of sickle cell anemia is described based on an allele-specific polymerase chain reaction (ASPCR). This method allows direct detection of the normal or the sickle cell beta-globin allele in genomic DNA without additional steps of probe hybridization, ligation, or restriction enzyme cleavage. Two allele-specific oligonucleotide primers, one specific for the sickle cell allele and one specific for the normal allele, together with another primer complementary to both alleles were used in the polymerase chain reaction with genomic DNA templates. The allele-specific primers differed from each other in their terminal 3' nucleotide. Under the proper annealing temperature and polymerase chain reaction conditions, these primers only directed amplification on their complementary allele. In a single blind study of DNA samples from 12 individuals, this method correctly and unambiguously allowed for the determination of the genotypes with no false negatives or positives. If ASPCR is able to discriminate all allelic variation (both transition and transversion mutations), this method has the potential to be a powerful approach for genetic disease diagnosis, carrier screening, HLA typing, human gene mapping, forensics, and paternity testing.
684 citations
TL;DR: The results suggest that the function of the 3' → 5' exonuclease activity of DNA polymerases is to remove mispaired nucleotides which have been incorrectly incorporated, thereby increasing the fidelity of template copying.
545 citations
TL;DR: The retrovirus that causes AIDS has revealed enough of its life history for a variety of therapeutic strategies to be apparent that are suitable for immediate application in clinical trials or have already yielded positive results in some patients.
Abstract: The retrovirus that causes AIDS has revealed enough of its life history for a variety of therapeutic strategies to be apparent. Some of these are suitable for immediate application in clinical trials or have already yielded positive results in some patients.
470 citations
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3,342 citations
TL;DR: In the studies to be reported here, the frequencies of the 16 possible nearest neighbor pairs in a variety of DNA’s are derived by the technique of enzymatic incorporation of 5'-Pazlabeled nucleotides into DNA and then degradation of the DNA into 3’-nucleotides.
673 citations
TL;DR: Bello, Van Bibber, and Bessman have shown that the deoxyguanylate kinase of infected cells is physically distinct and separable from the host cell enzyme, and the polymerase of uninfected E. coli (E. coli polymerase) and the T2 polymerase are distinct and different enzymes.
556 citations
420 citations