I would like to know what are the conventional methods to purify DNA and PUC19 after digestion with restriction enzyme.
i have used sodium acetate (pH 4.5) method but i have lost the bands when running gel electrophoresis.
RE used: Ecor1 and Kpn1
Hi there,
How sure the problem isn't at the digest level? Did you check the digest profile on gel before purification?
I have checked the bands after digestion... there were clear bands.
Hi again,
What do you call sodium acetate method? Is it just ethanol precipitation in presence of this salt? If yes then either the preciptation hasn't work or DNA got lost or degraded during the process.
I always go for DNA band extraction from agarose gel (even if yield is never great) and then purification on silica column using commercial gel extraction kit (Qiagen, Thermo, Macherey Nagel...)
Thank you. I used the same sodium acetate for pcr product purification. I got the bands. But the band lost after purification for digested product. is there any conventioanl method for purification. The kit which i purchase have some problem. That is why im using conventional way.
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