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Extraction of large size genomic DNA


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#1 Nrelo

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Posted 16 April 2009 - 01:12 AM

I want to extract genomic DNA which is up to several mega base pair from Burkholderia, are there any commercial kits capble to do so? Thanks

#2 phage434

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Posted 16 April 2009 - 04:08 AM

What do you want to do with this DNA. If you need it in whole chromosomes, then you need to embed cells in agarose and digest them in-place. If you just need DNA for sequencing, library construction, or PCR, then any normal genomic prep will work fine. I've had a lot of success with the Zymed 96 well plate format preps, but also with the Sambrook CTAB protocol.

#3 Nrelo

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Posted 16 April 2009 - 04:54 AM

What do you want to do with this DNA. If you need it in whole chromosomes, then you need to embed cells in agarose and digest them in-place. If you just need DNA for sequencing, library construction, or PCR, then any normal genomic prep will work fine. I've had a lot of success with the Zymed 96 well plate format preps, but also with the Sambrook CTAB protocol.


I have inserted a gene into the bacterial genome and want to find out the insertion site. So I need to extract the genomic DNA, digest it with a R.E, ligate the fragments and sequence them. In this case, any normal genomic prep will work?

#4 phage434

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Posted 16 April 2009 - 08:00 PM

Yes.




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