Suppression Subtractive Hybridization (SSH) query!

Csaba Kiss csakis at ki.se
Mon Jan 31 13:08:44 EST 2000


Hi!

    I am planning to perform SSH on my samples. I have not done the protocol
before, so I am not experienced with it. I read all the available info on
the method. It seems to me that it is a very powerfull method. I would
appreciate any tips and tricks from those who did this before, what to look
for, what to be really carefull with. (I.e. Ligating the Adaptors to the end
of the cDNBA fragments is one of the crucial steps.) Things like this,
hybridization time, temperature, PCR cycle number.
    I thought that I would buy the necessary components separately, but it
seems to me it is much easier (not much more expensive!) just to buy the kit
from Clontech. Have you used the kit? Did it work well?

Any suggestions are welcomed!

Thanx

**************************
       Csaba Kiss
      csakis at ki.se
      ICQ #7911383
 MTC Karolinska Institute
    Stockholm, Sweden
**************************






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