I performed a gel extraction of a PCR product and after elution at the last step, the eluted product becomes solidified. I use 0.4% gel for extraction. I do step wise elution of 20+20 ul after incubation at 37C for 5min. The kit used is Qiagen DNA extraction kit.
It sounds like some agarose gel is still getting through. I would recommend following the additional wash steps in the protocol. We routinely do gel extractions from 0.8% agarose gel and have no problems.
Michael Jaskolka Do you recommend doing second wash with Gel wash buffer provided? I generally use 0.4% to easily dissolve the gel. Will the percentage of agarose have an effect?
Santanu Sasidharan Yes, I would recommend doing the second wash. The protocol says to heat for 10min, but I would strongly recommend letting it go for longer. Heat until no schlieren is seen when mixing. The schlieren is unmelted agarose.
I have used different brands including Qiagen for gel extraction. But, I remember that all used 55 degC to 65 degC to melt gel blocks complete with intermittent votexing. The 37 degC that was used may not be enough to melt it completely.
@Taek H. You . Thank you for the reply sir but i have melted the gel blocks at 60C. But after eluting the product, the 20-30ul of the elution becomes solid again. This problem is hindering my further step in cloning.
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