The production of single-stranded DNA suitable for sequencing using the polymerase chain reaction |
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| Authors: | M W Allard D L Ellsworth R L Honeycutt |
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| Institution: | Department of Wildlife and Fisheries Sciences, Texas A&M University, College Station 77843-2258. |
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| 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. |
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