cDNA library screening - (Jan/17/2001 )
I was wondering about probe length in regards to library screening. I have a 100 bp rtPCR product from the coding region of a gene and I am wondering if it will be long enough to use as a radioactive probe to successfully screen a cDNA library. (The RNA used for the library and for the rtPCR is from the same organism at the same developmental stage) What is the minimum probe length anyone has had success with under these conditions? Thank you in advance for any insight.
Probe length is important when screening cDNA libraries. I would not recommend using a probe under 400 bp.We have had results using a probe of 250 bp but not under that. Basically the bigger the probe the better.