Production of single-stranded DNA for sequencing: an alternative approach.
We describe a simple procedure for the direct sequencing of single-stranded, PCR-amplified, target regions of human genomic DNA. At variance with previously reported procedures, purification of the desired double-stranded DNA was introduced. This additional step allowed the single-stranded amplification and sequencing of the target gene. This step is required for direct sequencing of some amplified regions of human genomic DNA. However, no individual technique seems suitable to generate and sequence all single-stranded DNA.
Medori
R
R
Division of Neuropathology, Case Western Reserve University, Cleveland, OH 44106.
Tritschler
H J
HJ
Gambetti
P
P
D001483:Base Sequence; D001709:Biotechnology; D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification*; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D018620:PrPSc Proteins; D011328:Prions / Q000235:genetics
D004277:DNA, Single-Stranded; D018620:PrPSc Proteins; D011328:Prions