Producing single-stranded DNA probes with the Taq DNA polymerase: a high yield protocol.
We report an efficient procedure to synthesize either single- or double-stranded probes labeled with biotin-11-dUTP, biotin-21-dUTP or digoxigenin-11-dUTP. To produce the single-stranded probe, only a single primer is utilized in a Taq polymerase amplification of 55 cycles. A cytomegalovirus probe is presented. This procedure allows easy production of nonradioactively labeled pure single-stranded probes of any desired length and specificity.
Finckh
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U
Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
Lingenfelter
P A
PA
Myerson
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D
D001483:Base Sequence; D015342:DNA Probes; D004277:DNA, Single-Stranded / Q000302:isolation & purification*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D019914:Taq Polymerase
D015342:DNA Probes; D004277:DNA, Single-Stranded; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase