Abstract
The acceptance of polymerase chain reaction (PCR) as a routine method in molecular biology has created a need for simple and robust approaches to DNA sequencing of the amplification products. In addition to its ease, direct sequencing of PCR products has the advantage that nucleotide misincorporation during amplification does not pose a problem, because only a small percentage of the DNA molecules may contain such random mutations. However, a large body of published protocols attests to the difficulties encountered in this endeavor. Double-stranded sequencing of the linear PCR products is notoriously unreliable because of rapid reannealing of the denatured DNA.
Keywords
- Polymerase Chain Reaction
- dNTP Concentration
- dsDNA Fragment
- Asymmetric Polymerase Chain Reaction
- ssDNA Sequencing
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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© 1996 Humana Press Inc., Totowa, NJ
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Kaltenboeck, B., Kousoulas, K.G. (1996). Efficient PCR Production of Single-Stranded DNA Sequencing Templates. In: Rapley, R. (eds) PCR Sequencing Protocols. Methods in Molecular Biology™, vol 65. Springer, Totowa, NJ. https://doi.org/10.1385/0-89603-344-9:149
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DOI: https://doi.org/10.1385/0-89603-344-9:149
Publisher Name: Springer, Totowa, NJ
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