Microarray-based subtractive hybridzation
AssigneeAventis Pharmaceuticals Inc.
InventorsSteven Perrin, Kathy Call, Timothy Connolly
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Abstract
The present invention relates to a highly efficient, high-throughput method for the identification and elimination of redundancy in a population of nucleic acid molecules using microarrays. This method involves a reiterative subtraction protocol that creates a library that becomes more biased toward unknown genes with each successive round. The removal of repetitive and previously characterized nucleic acids from the library allows the identification of low-abundance mRNA from sources of interest and enhances the rate of novel gene discovery. The present invention is also useful for the removal of contaminating nucleic acids from cloning libraries.
Claims5
A method of enhancing the rate of novel gene discovery in a population of previously-uncharacterized nucleic acid molecules, comprising
- a. contacting an first ordered array of nucleic acid samples from said population with an initial complex subtraction probe pool, said probe pool comprising a labeled probe corresponding to a redundant or known sequence;
- b. identifying a nucleic acid, said nucleic acid being characterized by weak or no hybridization to said initial complex subtraction probe pool;
- c. sequencing said nucleic acid to yield a newly-identified sequence;
- d. adding said newly-identified sequence to said initial pool to generate a second complex subtraction probe pool, said second pool comprising a larger number of labeled probes compared to said initial pool;
- e. contacting a second ordered a array of nucleic acid samples from said population with said second subtraction probe pool;
- and
- f. repeating steps b.-e., wherein each repeat of steps b.-e. reduces the redundancy of said population of previously-uncharacterized nucleic acid molecules, thereby enhancing the rate of novel gene discovery.
Dependencies are inferred from the claim text.