Design of a high-throughput assay for alternative splicing using polymerase colonies.
We propose an assay to detect and quantify alternative splicing simultaneously for numerous genes in a pool of cellular mRNA. The assay exploits polymerase colonies, a recently developed method for sampling and amplifying large numbers of individual transcript molecules into discrete spots on a gel. The proposed assay combines the advantages of microarrays for transcript quantitation with the sensitivity and precision of methods based on counting single transcript molecules. Given a collection of spots s(i), each containing an unknown splice variant of some known gene G(i), we design a series of hybridizations to short oligonucleotide probes to determine in parallel which exons of G(i) are present in every spot s(i). We give algorithms to minimize the cost of such designs.
Department of Computer Science and Engineering, Washington University, One Brookings Drive, St. Louis, MO 63130, USA. jbuhler@cse.wustl.edu
J D
JD
Buhler
R M
RM
Souvenir
W
W
Zhang
R D
RD
Mitra
D017398:Alternative Splicing; D019295:Computational Biology; D005091:Exons; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D005821:Genetic Techniques; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics
D012333:RNA, Messenger