Hi,
I need to produce around 100 µg of 24 different DNA fragments (In total 2.4 mg). The length of the fragments varies from 700 bp - 2000 bp. The fragments will later be used for digestion and ligation to assemble different plasmids and transformed into E. coli. My question is now how to I do this in a wise way?
My immediate proposal would be to amplify the fragments in 50 µL PCRs and subsequently purify the reactions. And here comes a second question: Do you know of any PCR purification kits that takes large volumes?
Vikas Kumar Patel
Dear Viktor,
You can do PCR reaction in multiple vials, pool them and load in a single fused agarose well.I would suggest you to use Qiagen gel elution kit. For amplification use hi-fidelity fusion polymerase.
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