ide 5'-triphosphates (dNTPs) were purchased from Pharmacia.
Oligonucleotide primers and probes were synthesized on
a Biosearch 8700 DNA synthesizer. Oligonucleotide probes
were synthesized with a 3'-phosphate to prevent extension
(7). [_y-32P]ATP (3000 Ci/mmol; 1 Ci = 37 GBq) was from
New England Nuclear. Single-stranded M13 DNA template
(M13mp10) was prepared as described (8). Human immunodeficiency
virus type 1 (HIV-1) DNA template (GeneAmplimer
HIV-1 positive control DNA, Perkin-Elmer/Cetus)
contained the HIVZ6 genome, which was rearranged to
interrupt the pol gene region and block infectivity, and cloned
into the plasmid pBR322 (9). Bacteriophage A DNA template
(GeneAmp positive control DNA) was obtained from PerkinElmer/Cetus.
Human genomic DNA was prepared from the
human cell line HL60 as described (10) and, where indicated,
was degraded by hydrodynamic shearing in a French press
(Aminco) to =800 base pairs (bp) in size.