(by Anne.Roessger from klinik.uni-regensburg.de)
Thu May 24 10:55:36 EST 2007
Thank you Peter for your help.
I´ll try to find the sequences next to my sensor then by RACE or
restriction-site PCR. I try to find the integration site of my used
transposon. If anybody does know another method to find the integration
sites it would be very helpful for me.
"peter" <peter.ianakiev from gmail.com> schrieb im Newsbeitrag
news:1180017336.890275.280240 from o5g2000hsb.googlegroups.com...
> On May 24, 2:50 am, "Anne" <Anne.Roess... from klinik.uni-regensburg.de>
>> By PCR? You cut the nylonmembrane and run a PCR with it or how do you do
>> "peter" <peter.ianak... from gmail.com> schrieb im
>> Newsbeitragnews:1179930831.833752.233410 from g4g2000hsf.googlegroups.com...
>> > On May 23, 8:29 am, "Anne" <Anne.Roess... from klinik.uni-regensburg.de>
>> > wrote:
>> >> Hello,
>> >> does anybody know if it is possible to get the DNA out of the
>> >> nylonmembrane
>> >> after crosslinking ? I have to know the sequences next to the binding
>> >> site
>> >> of my sensor.
>> >> Thanks a lot
>> >> Anne
>> > Yep, by PCR....
> That's right Anne - cut the membrane with your fragment on a small
> pieces and then use it in a PCR with your favorite primers. Just be
> careful to wash out all the SDS which will inhibit PCR reaction even
> at very low concentrations. Also for "crosslinking" might be better if
> you just use NaOH rather than UV, but that will not interfere with the
> PCR if your amplicon is small.
> Good Luck!
> PS. Sorry to the group for the multiple posts earlier - it is now
> obvious that I am using Windows PC....
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