U266 cDNA libraries cloning

Wolfgang Schechinger hubahopp at gmx.de
Tue Apr 29 02:57:54 EST 2003


starting with a library might be overkill: You have to make the library and
the bait and then fish for your gene without any warranty that you will be
able to isolate a complete clone.

But: you already know a lot about your target! It's not just a few amino
acids of a putatively new protein, the classical starting scenario for a
library screen. 

You can extract information from the GenBank or another database to
construct primers in order to catch at least some pieces of the cDNA. Then
perform primer walking, RACE or similar techniques to get the rest. 


At 05:08 29.04.2003 -0000, you wrote:
>Hi, all:
>I wonder if anyone could shed some light on the following question for me:
>I'm interested in cloning the complete cDNA of IgE secreted by the U266
cell line, but don't know how to get started.  Should I start by
constructing the entire cDNA libraries from U266?  Then, where can I find
the primers needed for singling out that IgE cDNA I need?  Any suggestion
will be highly appreciated, thanks.


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