Clontech PCR-Select subtractive Kit
chuiyl at hkstar.com
Sun Jan 26 03:56:12 EST 1997
Dr. B. Popping wrote:
> Dear Netters,
> last week I posted an article asking about your experience with the
> Clontech PCR-Select kit (suppresser subtractive hybridization).
> The only replies I had so far were people asking to forward any
> information I'd get to them.
> I can't quite believe that no-one has used the kit. Or is the experience
> you have made with the kit so BAD that you do not want to share it ?
> I'm still hoping to get information from poeple who have used the kit,
> even if they made had a negative experience.
> Bert Popping
> Bert Popping
> E-MAIL: bpopping at hgmp.mrc.ac.uk
Dear Dr. Popping,
Your message has just caught my attention. I am sorry that you haven't received response from this
newsgroup regarding your PCR-Select matter.
As a matter of fact, I happen to be one of those who is using this PCR-Select kit from Clontech. We
are attempting to identify genes which are differentially expressed in cancer cells. Our experience
so far in using the kit is still quite limited. We have gone through the PCR-Select procedure twice;
our impression is that the kit is user-friendly. The kit provides protocols for checking the progress all
along the entire procedure. We find that our results match well with their "standard". Using the
semiquantitative PCR method suggested by the manufacturer, we note that our subtracted
samples did have the amount of a housekeeping gene cDNA, G3PDH, reduced to a level
comparable the one shown in the manual. We are now in the process of doing Southern of the
cDNA to find out more accurately to what extent the G3PDH cDNA was subtracted. We are also
using the subtracted cDNA as probe for Northern blotting to find out how good is the enrichment. I
shall keep you informed of our result when it is known. I am sorry that we are not yet in a position to
tell you more.
I do have a colleague working in our neighbouring lab. His name is Dr. John Chan; he managed to
use the PCR-Select kit to isolate a gene, I think, related to breast cancer. The work was published
in PNAS. I wonder is that paper the one you have. I shall give you detail on this if they are not the
Yiu-Loon Chui, Ph.D
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