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稀巴烂

银虫 (初入文坛)

[求助] 反硝化功能基因反应条件摸索~~~ 已有1人参与

如题,摸索了好长一段时间的反应条件,在反硝化过程中从NO3 -还原成NO2 -的过程中,很多文献都是用narG-1960F(TAYGTSGGSCARGARAA) & narG-2650R(TTYTCRTACCABGTBGC),反应条件为:95℃-5min;95℃-30s/59 to 55℃-30s/72℃-45s(前八个循环每一个循环-0.5℃,后30cycles保持55℃);72℃-10min。退火温度也换了几次,但是因为是递减pcr,又不能做梯度pcr,换了好几种酶(Ex Taq、HS Taq、MightAmp DNA Polymerase、加BSA也没用),这功能基因不知道有哪位虫友做过呢?谢谢啦~
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迷路小小书童

新虫 (初入文坛)

【答案】应助回帖

Detection of genes for membrane-bound nitrate reductase in nitrate-respiring bacteria and in community DNA
你可以看看这篇文献,考虑用W9 T38 作为引物
我最近也要做这个基因PCR
2楼2014-07-21 15:12:13
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