DNA degradation in BAC preps - DNase inhibitor? (Apr/12/2007 )

I agree that your preparation of DNA is likely to be the problem. How are you doing it?

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Sigma NA100 kit.

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Hi, I also found that my BAC preps degraded quite quickly using this Kit. It was much better if I missed out the endotoxin removal step, and I then went on to electroporate this DNA into ES Cells.

I now use another Kit from Macherey- Nagel (Nucleobond BAC 100) and find this is much better.

Good Luck

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Thanks, I had some fears about the kit. I'll try it without the endo step. I actually tried t get a quote for the nucleobond kit but they never got back to me! Do you mind me asking how much it cost? (I'm in the UK)

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I got my Kit from Abgene, £176.00 for 10 preps Cat No MN740579, but that was about a year ago, so it may well be a bit more.

If you want the method I use which is slightly different to the booklet they provide, I will send it to you. Just let me know.

Where are you based in the UK?

Cheers

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I'm at Liverpool uni. Are you UK too?

PS. I'd really appreciate that protocol! Thanks

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Yip I am in Edinburgh. I will send the method as an attachment to your PM.

Probably tomorrow now as I am not at my usual computer.

Ok

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