HELP HELP cDNA library

[Chi-kuang Wen]

Els Van Geldre (Els.VanGeldre at rug.ac.be) wrote:
: Hi,

: I synthetised cDNA starting from mRNA from plant sources. After addition 
: of Eco RI/Not I adaptors and insertion into lgt 11 phage I only become 
: blue plaques. The problem is probably the addition of the 
: EcoRI/Not I adaptors or the insertion into the lgt 11 phage. However, I 
: strictly followed the instructions of the kit I used (TimeSaver cDNA 
: synthesis kit). Do you have any suggestions to become recombinant plaques 
: ?

: Thanks,
: Els Van Geldre
Hi, Els: 
  Did you size-fractionate the insert/adaptor ligation rxn?  If the adaptor is not removed, it will self-ligates and becomes inserts to generate blue phages.

Chi-Kuang