I have spent nearly a year now trying to clone the 5' end
of a particularly stubborn cDNA. I have tried rescreening the
library (and other libraries) with a restriction fragment on
the 5' side of my original (incomplete) clone and all the
clones (12 of them) were shorter than the original.
Lately I have tried the clontech 5' amplifinder kit which uses
RNA ligation to anchor the 5' end. In short this was a miserable
failure ..not even the control that came with the kit worked.
I have called clontech and they say they have "had problems with it"
So I have two questions
1) Those of you who have succeeded in getting your 5' end.
What do you think is the best approach?
and secondly and less importantly
Do clontech suck?
Any and all answers (esp to the first question) will be much appreciated.