clonetech cDNA construction kit
s.bhattacharya at hosp.wisc.edu
Thu Sep 4 11:32:48 EST 1997
Please consider this as an SOS signal:
I have been trying to pull out a full length clone from Clonetech's smart cDNA
construction kit. After I went through all the pain of constructing the library,
primary to secondary to tertiary to quarternary screens by filter lifts etc I found
a partial clone (500 bp) nowhere close to full length. My isolated RNA is good
and my Northern data shows it is a 2.5 kb message.
This is very upsetting for me because I have no confidence on this kit
anymore,because if their kit (smart oligo) was working I should only get full
length clone, or I donot get any clone. They claim their smart oligo should only
bind to 5' end of any message.
If you know of someone who have used this kit succesfully would you please let
me know? My address is s.bhattacharya at hosp.wisc.edu
I would also welcome any idea to get this fulllength clone.
If you are wondering why I went to make my own library the answer is I donot
have this clone in a commercially available Jurkat Cell library , and I figured
out that making my own library would be easier than going through a no of
differnt libraries from the market.
Thank you for your time.
More information about the Methods