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• W1986665555 endingPage "5033" @default.
• W1986665555 startingPage "5026" @default.
• W1986665555 abstract "We characterized the behavior of polymerase chain reactions (PCR) using degraded DNA as a template. We first demonstrated that fragments larger than the initial template fragments can be amplified if overlapping fragments are allowed to anneal and extend prior to routine PCR. Amplification products increase when degraded genomic DNA is pretreated by polymerization in the absence of specific primers. Secondly, we measured nucleotide uptake as a function of template DNA degradation. dNTP incorporation initially increases with increasing DNA fragmentation and then declines when the DNA becomes highly degraded. We demonstrated that dNTP uptake continues for >10 polymerization cycles and is affected by the quality and quantity of template DNA and by the amount of substrate dNTP. These results suggest that although reconstruction of degraded DNA may allow amplification of large fragments, reconstructive polymerization and amplification polymerization may compete. This was confirmed in PCR where the addition of degraded DNA reduced the resultant product. Because terminal deoxynucleotidyl transferase activity of Taq polymerase may inhibit 3' annealing and restrict the length of template reconstruction, we suggest modified PCR techniques which separate reconstructive and amplification polymerization reactions." @default.
• W1986665555 created "2016-06-24" @default.
• W1986665555 creator A5007544675 @default.
• W1986665555 creator A5055479254 @default.
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• W1986665555 date "1996-12-01" @default.
• W1986665555 modified "2023-09-29" @default.
• W1986665555 title "Effect of Highly Fragmented DNA on PCR" @default.
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• W1986665555 doi "https://doi.org/10.1093/nar/24.24.5026" @default.
• W1986665555 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/146324" @default.
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• W1986665555 hasPublicationYear "1996" @default.
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