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-   -   Do proteins interfere with NGS or lib prep?? (http://seqanswers.com/forums/showthread.php?t=11083)

hmviit 05-02-2011 11:29 PM

Do proteins interfere with NGS or lib prep??
 
Hi all,

I found this forum a week a ago and it has turned out to be useful. The only thing I'm still wondering about is said in the title. I will be doing DNA sequencing on 454 after seq capturing. I suspect that the gDNA that I have after extraction has RNA as a contamination (liver tissue so I expect RNA) and I want to treat the gDNA with RNAse A. But after that I'm not sure how important it is to get rid of the RNAse in the gDNA and by serching around I haven't found a good answer. I'm using salt-extraction and dissolving my gDNA in to H2O. So after treating with RNAse, should I re-extract or just precipitate to get rid of the RNAse-proteins or just let it be?

Thanks,
Heidi

gogreen 05-08-2011 07:05 AM

Hi Heidi, I would recommend a cleanup of gDNA (columns, beads or phenol ChCl3) before you do library prep. If I got it right, you want to use the DNA for sequence capture before actual sequencing. Having clean DNA in first place would already help a lot in the downstream.


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