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et2b's Content

There have been 3 items by et2b (Search limited from 17-November 18)


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#174829 DNA yields after Bisulfite Treatment

Posted by et2b on 13 May 2015 - 01:52 AM in DNA Methylation and Epigenetics

On the 260/280 topic. Bisulfite conversion does not only make your DNA single stranded, but also degrades the DNA. This results in many small fragments with an -OH group at the end, which will thus "pollute" your spectro profile in the range of the ethanol group, so that you 260/280 is no longer 'clean' . 

 

@ people doing double stranded DNA quantification after bisulfite conversion: do a single stranded method and your yield will be better :-). Bisulfite conversion yields non-complementary DNA strands.

 

best wishes




#174827 Size selection beads

Posted by et2b on 13 May 2015 - 01:46 AM in Molecular Biology

http://core-genomics...beads-work.html

best wishes




#174825 bisulfte conversion with Ampure XP beads in the solution

Posted by et2b on 13 May 2015 - 01:40 AM in DNA Methylation and Epigenetics

Dear all,

 

I wonder if anybody ever tried to do bisulfite conversion with Ampure XP beads (SPRI) in the solution?

I know zymo research has a magbeads protocol. But according to zymo their beads have no size selection properties.

 

What I would like to do is a protocol similar that of Fisher et al. http://genomebiology...content/12/1/R1

keeping all steps on Ampure XP beads...  (have very very little DNA...)

So fragmentation or restriction of gDNA, end-repair, adapter ligation, size selection of >150bp, bisulfite conversion, cleanup, desulfanation,cleanup 

 

best





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