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• W2062296959 abstract "Primer extension analysis was evaluated as a means to identify PCR-amplified DNA from HIV-1. Solution hybridization with radioactive labeled oligonucleotides followed by an extension reaction with Klenow fragment of Escherichia coli DNA polymerase I and subsequent separation on denaturing polyacrylamide gels reveals single stranded DNA products of the predicted size. The specificity of the reaction is further demonstrated by specific endonuclease digestion. The analysis is more sensitive than Southern blotting and about equally sensitive as Slot blot analysis when double-stranded DNA probes are used for hybridization. With end-labeled oligonucleotide probes, primer extension analysis proved an order of magnitude more sensitive than membrane hybridization. The analysis also allows quantitation of amplified DNA from 1 pg to about 1 ng of DNA product. Under the conditions described for amplification, primer extension analysis is capable of detecting a single HIV-1 plasmid DNA molecule in the presence of 1 microgram of total DNA. 3'-end mismatching of the oligonucleotide probe does not result in a significantly altered detection limit. Primer extension analysis can also be carried out with at least three different PCR-amplified DNAs in the same reaction tube." @default.
• W2062296959 created "2016-06-24" @default.
• W2062296959 creator A5001947909 @default.
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• W2062296959 date "1993-05-01" @default.
• W2062296959 modified "2023-10-14" @default.
• W2062296959 title "Primer extension analysis provides a sensitive tool for the identification of PCR-amplified DNA from HIV-1" @default.
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• W2062296959 doi "https://doi.org/10.1016/0166-0934(93)90042-p" @default.
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