An improved method for constructing a full-length enriched cDNA library using small amounts of total RNA as a starting material

We have developed an improved method for con-structing a ful-length cDNA library using smal quantity of material by modifying the original oligo-capping method. In our devised method, total RNAs are used in sequential oligo-caping steps directly without preliminary mRNA puri-fication. Using this method, we constructed ful- length cDNA libraries from 10 g of total RNA. These libraries contained 8105 to 8106 inde-pendent clones with average insert sizes of 2.0 kb. Moreover, the number of ful-length cDNAs con-taining the translation initiation codon ATG in the constructed libraries was estimated to 60-70%. In adition, 54% of the known cDNAs had a longer 5' end than the coresponding genes in the public database. Our results show that the method can be effectively used to construct ful-length enri-ched cDNA libraries, especialy, if starting material is limited. KCI Citation Count: 15