APPLICATION

Rapid-Screen™ Arrayed cDNA Libraries can be used to clone alternatively spliced transcripts, even if the desired spliced variant is many times less abundant than its parent transcript. The detection of different-sized fragments through use of the 5' vector primer plus the 3' gene-specific primer is sometimes attributable to alternative-splicing rather than to truncated transcripts. If the alternate splicing has already been characterized, gene-specific primers that differentiate between the two spliced forms can be designed.

Rapid-Screen Arrayed cDNA Libraries were used with a pair of gene-specific primers to demonstrate a pooling technique for the detection of two alternatively spliced transcripts of a gene. The shorter form was much more abundant than the longer one. The shorter appeared in seven of the twelve column pools (40,000 clones/pool) and four of the eight row pools (60,000 clones/pool). The longer form (approximately 210 bp longer) was found only in the column 1 pool and in the row H pool. Thus well H1 of the Master Plate contains the lower abundant transcript variant.

RFAL Fig 1