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魏茂繁新蟲(chóng) (初入文壇)
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[求助]
PCR
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| 我的引物GC含量過(guò)高,Tm溫度過(guò)高達(dá)到75度,那我該怎么辦 |
新蟲(chóng) (初入文壇)
木蟲(chóng) (小有名氣)
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直接用72攝氏度退火延伸進(jìn)行實(shí)驗(yàn),也可能可以擴(kuò)增出來(lái) PCR體系加入 DMSO有利于高GC的擴(kuò)增 DMSO is used in PCR to inhibit secondary structures in the DNA template or the DNA primers. It is added to the PCR mix before reacting, where it interferes with the self-complementarity of the DNA, preventing the occurrence of interfering reactions. However, use of DMSO in PCR increases the mutation rate. PCR體系中DMSO的量一般不超過(guò)總體積的4%。 |
新蟲(chóng) (初入文壇)
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