Published online 15 December 2004
Nucleic Acids Research, Vol. 32 No. 22 © Oxford University Press 2004; all rights reserved
Evaluation of vector-primed cDNA library production from microgram quantities of total RNA
Laboratory of Genetics, National Institute of Mental Health Building 35, Room 1B-215, 35 Convent Drive, Bethesda, MD 20892-3728, USA
* To whom correspondence should be addressed. Tel: +1 301 402 6976; Fax: +1 301 402 6976; Email: usdint{at}mail.nih.gov
Received October 6, 2004; Revised November 16, 2004; Accepted November 29, 2004
cDNA sequences are important for defining the coding region of genes, and full-length cDNA clones have proven to be useful for investigation of the function of gene products. We produced cDNA libraries containing 3.5–5 x 105 primary transformants, starting with 5 µg of total RNA prepared from mouse pituitary, adrenal, thymus, and pineal tissue, using a vector-primed cDNA synthesis method. Of 
1000 clones sequenced, 20% contained the full open reading frames (ORFs) of known transcripts, based on the presence of the initiating methionine residue codon. The libraries were complex, with 94, 91, 83 and 55% of the clones from the thymus, adrenal, pineal and pituitary libraries, respectively, represented only once. Twenty-five full-length clones, not yet represented in the Mammalian Gene Collection, were identified. Thus, we have produced useful cDNA libraries for the isolation of full-length cDNA clones that are not yet available in the public domain, and demonstrated the utility of a simple method for making high-quality libraries from small amounts of starting material.