I just did this and I read that vortexing 10 kb + DNA fragments is not recommended. I am wondering if I lost my plasmid. Ultimately, I would like to ligate an insert into this plasmid. They were previously digested and they were vortexed as part of the gel purification.
Your linear DNAs should be fine. Vortexing is normally part of the gel extraction/purification protocol. Although I do recommend running a small amount of your purified DNA on a gel to be sure you recovered it and get an approximate concentration. It will also show if the DNA is intact or not.
Good Luck!
Dear Sir,
Yes it is correct that vortexing is not recommended for larger fragment. But as per kit protocol for Gel purification/extraction vortexing is not required. You have to just melt the gel in extraction buffer and tilt it 4-5 times. Then pass it through the column and elute the DNA fragment.
I think that for future it would be wise not to vortex, but I highly doubt it will cause you any issue with this cloning work. In general, DNA can break when vortex ing down to 10-12kb pieces, so once you are using plasmid in that size range and larger you definitely increase the chance of some breakage. You should be safe at 9kb.
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