Related Experiment Videos
The production of single-stranded DNA suitable for sequencing using the polymerase chain reaction
M W Allard1, D L Ellsworth, R L Honeycutt
1Department of Wildlife and Fisheries Sciences, Texas A&M University, College Station 77843-2258.
Abstract:
A simple and reliable procedure for the amplification of single-stranded DNA suitable for sequencing is described. This procedure employs the polymerase chain reaction and implements modifications pertaining to the purification of the double-stranded DNA product prior to single-stranded DNA amplification. The most consistent sequencing reactions are obtained when the double-stranded DNA product is purified by centrifugation with a microconcentrator prior to single-stranded DNA amplification and the overall amount of specific primers and number of cycles used, in both single-stranded and double-stranded DNA polymerase chain reactions, are reduced.