Cloning Bisulphite modified

Jim Hughes jhughes at pinnacle.jr2.ox.ac.uk
Tue Nov 6 10:32:40 EST 2001


Hi

has anybody noticed bias in cloning PCR products generated from bisulphite
modified genomic.  A collegue who is using this to assay the methyation
status of a CpG island he is interested in seems only to get the
unmethlyated form, even though the other form is readily seen by direct
sequencing and Bisulphite SSCA.  He using pGEMt-easy from promega to do the
cloning.  Any thoughts?

Jim




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