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Gel Extracting a 15kb Band?


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#1 amisra2

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Posted 15 November 2010 - 03:16 PM

Hi

I am trying to extract a 15kb Band from agarose gel for ligation. I have used the Qiaquick Gel Extraction Kit, and it didn't work. The kit says it is designed to extract and purify DNA of 70bp to 10kB so I am not surprised. But is there another kit or another method out there that's easy, quick and cheap for larger fragments?

Any help would be much appreciated! Thank you

#2 bob1

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Posted 15 November 2010 - 05:06 PM

There's plenty of methods out there for ligation in the gel. Otherwise you could try eluting onto DE81 paper (see Sambrook et al Molecular cloning: a laboratory manual), or "freeze 'n' squeeze" (freezing the gel, spinning out the agarose and using the supernatant).

#3 perneseblue

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Posted 15 November 2010 - 08:15 PM

and you can try electrolution or beta agarase treatment.
May your PCR products be long, your protocols short and your boss on holiday

#4 BioMiha

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Posted 15 November 2010 - 11:34 PM

Or you could just use a genomic DNA extraction kit. It's designed for long DNA fragments.




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