Other Species Collections
pBluescriptR Library Method Overview
Dr. Michael Brownstein, National Institute of
Mental Health, NIH, Bethesda, MD and by Drs. Shiraki Toshiyuki and Piero Carninci (RIKEN) constructed
libraries in the vector pBluescriptR. The protocols used were based on the papers referenced below. Briefly, normalized and subtracted
cDNA libraries were prepared in a single step, to increase
the number of different cDNAs. This method:
- Was based on hybridization of the first-strand, full-length cDNA with several RNA drivers
- starting mRNA as the normalizing driver
- run-off transcripts from
mini-libraries containing highly expressed genes, rearrayed clones, and previously
sequenced cDNAs as subtracting drivers
- Kept the proportion of full-length
cDNAs in the subtracted/normalized library high.
- Enhanced the discovery
of new genes as compared to results obtained by using standard,
full-length cDNA libraries.
- Margolin J. Of Mice, Men, and the Genome.
Genome Res. 2000 Oct;10(10):1431-2.
- Carninci P, Shibata Y, Hayatsu N, Itoh M, Shiraki T, Hirozane T, Watahiki A, Shibata K, Konno H, Muramatsu M, Hayashizaki Y. Balanced-size and long-size cloning of full-length, cap-trapped cDNAs into vectors of the novel lambda-FLC family allows enhanced gene discovery rate and functional analysis.
Genomics 2001 Sep;77(1/2):79-90.
- Shibata Y, Carninci P, Watahiki A, Shiraki T, Konno H, Muramatsu M, Hayashizaki Y.
Cloning full-length, cap-trapper-selected cDNAs by using the single-strand linker ligation method.
Biotechniques 2001 Jun;30(6):1250-4.