The production of single-stranded DNA suitable for sequencing using the polymerase chain reaction.
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.
Allard
M W
MW
Department of Wildlife and Fisheries Sciences, Texas A&M University, College Station 77843-2258.
Ellsworth
D L
DL
Honeycutt
R L
RL
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D004277:DNA, Single-Stranded / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*
D004277:DNA, Single-Stranded