Sequencing of double-stranded PCR products.
Very often the experimental step following PCR is sequencing of the amplified fragment. Two protocols that allow direct sequencing of a double-stranded PCR product are described. The first involves removal of one strand of the PCR product using an M13 single-stranded DNA clone, allowing the second strand to be sequenced. The second protocol involves Maxam-Gilbert chemical sequencing after PCR amplification with one labeled primer. The advantages and disadvantages of the two protocols are compared, but both yield DNA sequence without cloning of the PCR product.
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Department of Biological Sciences, State University of New York at Binghamton, 13902-6000, USA.
D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*