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02:25 min
May 17th, 2024
DOI :
10.3791/201691-v
Transcript
为了执行基于CRISPR的模块化组装或CRISPRmass的平行两步试管反应,继续在试管反应步骤1中通过Cas9 sgRNA切割cDNA / ORF质粒的相同载体骨架。为此,将正确标记的 0.2 毫升管放置在冰上的铝制冷却块中。通过混合适量的Cas9,sgRNA,Cas9缓冲液和DEPC处理过的水,制备N次反应的预混液。
将预混液的组分加在一起后,涡旋相同的成分以确保正确混合。然后将混合物旋转并分装两微升预混液到每个 0.2 毫升管中。接下来,向每个试管中加入 0.4 微升 0
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