Cloning Bisulphite modified

Hans-Helmut Niller Hans-Helmut.Niller at klinik.uni-regensburg.de
Wed Nov 7 03:43:27 EST 2001


The only reliable way to determine CpG methylation state seems to be 
direct sequencing of bisulfite treated DNA.

Best,
Hans


>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

----------------------------------------------------
----------------------------------------------------
Dr. Hans Helmut Niller
Institut fuer Medizinische Mikrobiologie und Hygiene
Universitaet Regensburg
Franz-Josef-Strauss-Allee 11
D - 93053 Regensburg
Germany
phone:  +49 941 944-6451
fax:    +49 941 944-6402
e-mail: Hans-Helmut.Niller at klinik.uni-regensburg.de

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