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gzdeng铁杆木虫 (著名写手)
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由于没有生物背景，现需要翻译下面内容，怕翻译不准，请兄弟姐妹们帮帮忙，非常感谢！ DNA templates were prepared by the freeze-thaw method in a DNA extraction buffer(25). PCR was conducted for 40 cycles, with denaturation at 94°C for 60 s, annealing for 30 s, and extension at 72°C for 30s using the PROGRAM TEMP CONTROL SYSTEM (PC-800, ASTEC, Fukuoka). Here, considering that each Cl20 or C230 target sequence has a different homology score with the designed primers, a step down method was employed (26). For the Cl20 genes, the annealing temperature was 61’C in the first 10 cycles followed by a step down to 59°C in the next 15 cycles, and 57°C in the last 15 cycles, while for the C230 genes, the annealing temperature was 59°C in the first 10 cycles, 57°C in the next 15 cycles, and 55°C in the last 15 cycles. Aliquots (10~1) of the PCR products were analyzed by electrophoresis on a 1.0% agarose gel stained with 0.5 pg/ml ethidium bromide. |
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