IMPROVED METHOD FOR OBTAINING FULL-LENGTH cDNA SEQUENCES.

摘要

A method for obtaining longer cDNA sequences is provided. The method utilizes a known genomic DNA sequence or a partial cDNA sequence, such as can be obtained from GenBank partial cDNAs. Two PCR primers are designed to correspond to the ends of the known partial sequence and to anneal to DNA in a cDNA library so as to initiate extension away from the known cDNA and the other primer. The primers are added to a cDNA library with appropriate enzymes and extend through additional DNA sequence to produce PCR products, which are subsequently purified and sequenced to provide new sequences. The new sequences are then compared with the known partial cDNA sequence for areas of overlap, and the sequence is extended beyond the overlapping areas to provide longer DNA sequence.