Direct sequencing of long polymerase chain reaction fragments.
Direct sequencing of polymerase chain reaction (PCR)-generated templates is a commonly used technique in molecular biology laboratories. We describe an improved method for direct sequencing of PCR fragments longer than 20 kb obtained with a commercial mixture of Taq and Pwo DNA polymerases. The sequencing protocol was optimized for an automated infrared DNA sequencer, consistently yielding long reads (500-600 bases).
Iannelli
F
F
Dipartimento di Biologia Molecolare, Università di Siena, Italy.
Giunti
L
L
Pozzi
G
G
D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*; D013296:Streptococcus pneumoniae / Q000737:chemistry / Q000235:genetics
D004269:DNA, Bacterial; D004247:DNA