2.5　Detection and Quantitation of PCR Products

The PCR products were detected by solution oligomer hybridization.Ten microliters of PCR product and 3.3 μl of32P-labeled SK19 probe（（3-5）×105 cpm；3,000 Ci/mmol/L）in 10 mmol/L EDTA，15 mmol/L NaCl were heated to 95℃for 5 min，then hybridized at 55℃for 15 min.Five microliters of a dye containing 0.0l%Bromophenol blue，0.01%Xylene cyanole FF，50%glycerol，0.125 mol/L EDTA，pH 8.0 and 0.1%SDS was added.Aliquots of 5 μl were loaded on a 12%polyacrylamide minigel and run at 300 V for 30 min.The gel was exposed to X-ray film with intensifying screens at-80℃for 0.5 or 1 h.The bands on the gel corresponding to the HIV-1 gag sequence delineated by SK38/SK39（115 bp）were quantitated either directly with an Ambis Radioanalytic Imaging System（expressed as counts）or by cutting and counting the bands in a scintillation counter（expressed as cpm）.(https://www.daowen.com)