I am isolating PCR fragments from agarose gels slices. After preparing saturated NaI in water, I have found that after just a day the solution turns amber. Every day after that, the solution turns more and more amber and the performance of the solution to dissolve agarose drops. Is there a way to stabilize the saturated NaI, or must I make it fresh each day? Thank you in advance for any help on this.
Dear Steve
its been many years since i used gene clean top purify PCR amplicons and I remember the issue with discolouration of 6M NaI
If you read the pdf below:
- The oxidation culminating in yellow colour does not affect performance unless pH becomes alkaline
- Thus as suggested add 1/200 10% (v/v) glacial acetic acid or 1/30 TBE modifier
- It also suggests aliquoting out the stock solution. I would do this into opaque brown 2ml screw cap tubes to minimise photo oxidation
http://kirschner.med.harvard.edu/files/protocols/MPBIO_geneclean2.pdf
1/200 10% (v/v) glacial acetic acid or 1/30 TBE can use
http://kirschner.med.harvard.edu/files/protocols/MPBIO_geneclean2.pdf
Thanks all,
In case anyone is interested, a solution of guanidinium isothicyanate works best to dissolve agarose gel slices: much better than NaI.
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