Purification of Small DNA less than 100 bp Using Kit?
i have an issue with the Qiagen kits. My DNA is about 60 bp. Now its a bit too big for annealing two primers and a bit small to purify using the DNA kits (less than 100 bp a lot is lost)!
what can i do to purify this small dna!? :spcartmanmad:
thanks in advance
Re: Purification of Small DNA less than 100 bp Using Kit?
I used to do PCR diagnostics and we avoided kits for this same reason, (the 100bp cutoff). The very reliable method we used was DNA extraction via homemade reagents.
Briefly Lysis buffer, Pro-K trtmnt, boiling--and then either a) Chromaspin-30 column and ready for PCR or b) NaAcetate, Isopropanol precipitation (1 hour -20C incubation), EtOH washes, drying, resuspension, ready for PCR. As a footnote, if your target DNA is extremely small (less than 25bp) you can use LiCl instead of NaAcetate.
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