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Questions related from Ani Ben
Hello. Recently I've been assembling plasmids that use pSMPUW as the backbone (kanamycin resistant). I've noticed that the minipreps for these plasmids usually sequence fine (I've been using stbl3...
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I use T4 ligase buffer for Golden Gate because it doesn't contain PEG. However, I don't know its compatibility with BpiI. I was wondering if anyone has experience with this?
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I have a vector containing the EF1a promoter. I was wondering if DH5a cells will accept this vector upon transformation? I know some bacteria are incompatible with certain gene segments.
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Hello. So for a current project I'm doing, I'm assembling several fragments (6-7) using golden gate assembly. I've combined the ligation and digestion in to one reaction, which takes ~5 hours in...
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Say you have three fragments that have overlaps with each other. During PCR, I'm assuming that since these fragments could physically overlap with each other they will anneal. My question is will...
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