capture PCR

ELADER at AMBION.COM ELADER at AMBION.COM
Mon Sep 14 21:28:33 EST 1998


In article <v01530500b221b1a74393@[137.99.71.147]>,
  eul94001 at UCONNVM.UCONN.EDU (Eunkyung Lee) wrote:
> Dear colleagues,
>
> Did anyone have tried capture PCR using biotinylated primer and
> streptavidin beads? Can i have a protocol including which company products?
> What are the critical steps? Thanks in advance.
>
>

I did this with promega magnetic strep beads as a first step in a selection
process. This worked well, but the PCR step that followed made up for
efficiency.

I also used pipetman-tip-sized column called Affinitip (genosys sells them).
With a biotinylated pcr product (biotin on one primer) I could get nearly
quantitative yield of melting off the non biotinylated strand with 0.2N NaOH.

Hope this helps

Eric
elader at ambion.com

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