MBD-seq with bisulfite conversion

[lterhune]

Hi all,

Has anyone tried MBD-seq followed by bisulfite conversion yet? We are attempting MBD-seq first (MethylMiner) followed by a paired-end library and bisulfite conversion of the ligation before PCR. The limiting factor seems to be the input DNA quantity, since there is a ton of loss in the MethylMiner and bisulfite conversion steps.

Just wondering if anyone has been successful in this yet, and if so what input DNA/other parameters you used. Thanks!

[ECO]

Moving to sample prep

[frozenlyse]

http://genome.cshlp.org/content/earl....full.pdf+html was just published, where they performed MethylCap-bisulfite-seq.

[lterhune]

Thanks for the paper frozenlyse. We got DNA back at the end of our MethylMiner-PE library-BS conversion (about 0.3-1ng/ul) but aren't sure if it's enough to sequence.