The first goal was to develop methods that allowed generation of full length cDNA libraries. Reverse transcriptase protocols at the time had difficulties with the secondary structure of mRNA, leading to abbreviated cDNAs that were difficult to align and invited further complications in downstream analysis. To surpass this limitation, a method utilizing trehalose was developed to allow reverse transcriptase to function at a higher temperature, relaxing secondary structures. Other methods were additionally developed to assist in the construction of clonal cDNA libraries. These include a biotin-based capture system to select for full length cDNA, a novel lambda phage vector that minimized biases when delivering cDNA into a plasmid, and an iterative strategy to enrich for cDNA that had yet to be sequenced.
