SuperScript™ Pre-made cDNA Library, human bone marrow, mixed population

SuperScript™ Pre-made cDNA Libraries are suitable for isolating novel and published cDNAs, PCR of target sequences, and gene expression in eukaryotic cells (1). Each library is constructed using SuperScript™ II reverse transcriptase for first-strand synthesis to generate full-length and high-yield cDNA. The libraries have a large average insert size, typically 1.3 to 2.5 kb, ensuring high representation of full-length clones. Each library has greater than 5 x 10⁹ cfu derived from >10⁷ primary clones to ensure complete representation of even the rarest sequences. Libraries are amplified only once using a semi-solid amplification process for faithful representation of the primary library (2). Libraries are directionally cloned into the eukaryotic expression vector pCMV•SPORT containing a CMV promoter for expression analysis, an f1 ori for single-strand DNA production, and SP6 and T7 RNA polymerase promoter sites flanking the MCS for RNA synthesis. Libraries may be screened using conventional PCR or plate screening procedures, or by functional analysis using the eukaryotic CMV promoter.

Some of the SuperScript™ Pre-made cDNA Libraries are constructed in the pCMV•SPORT6 vector with Gateway™ Technology. Gateway™ is a universal cloning technology that provides a highly efficient and rapid route to functional analysis, protein expression, and the cloning/subcloning of DNA segments.

Please refer to Table 3 for detailed pCMV-SPORT vector information

Performance and Quality Testing: Qualified by PCR size analysis of 23 randomly selected clones to determine that 20/23 colonies contain inserts; average insert size is 1.2 to 2.5 kb. Animal libraries are screened for species-specific genes.

Contents and Storage:
Each library is supplied in 80% S.O.B., 20% (v/v) glycerol. Store at -70°C.

Reference(s):
1. Li, W. et al. (1995) Focus™ 17:45.
2. Kriegler, M. (1990) Gene Transfer and Expression: A Laboratory Manual. Stockton Press, New York, NY.