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METHODS, COMPOSITIONS, AND KITS FOR GENERATING rRNA-DEPLETED SAMPLES OR ISOLATING rRNA FROM SAMPLES

  • US 20110040081A1
  • Filed: 08/13/2010
  • Published: 02/17/2011
  • Est. Priority Date: 08/14/2009
  • Status: Abandoned Application
First Claim
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1. A method of generating a rRNA-depleted sample from an initial sample comprising:

  • a) providing;

    i) an initial sample comprising RNA molecules derived from at least one eukaryotic organism or species, wherein said RNA molecules comprise rRNA molecules and non-rRNA RNA molecules;

    ii) a composition comprising antisense rRNA molecules complementary to substantially all of the sequence of at least one rRNA molecule selected from the group consisting of;

    25S, 26S, 28S, 18S, 5.8S, and 5S eukaryotic cytoplasmic rRNA molecules and 12S and 16S eukaryotic mitochondrial rRNA molecules, wherein said antisense rRNA molecules comprise affinity-tags at a ratio of at least one affinity-tag per every 10 nucleobases of said antisense rRNA molecules; and

    iii) a binding matrix comprising affinity-tag-binding molecules;

    b) contacting said initial sample with said composition comprising antisense rRNA molecules under conditions such that at least some of said affinity-tagged antisense rRNA molecules and at least some of said rRNA molecules form double-stranded rRNA hybrids, thereby generating a treated sample;

    c) contacting said treated sample with said binding matrix under conditions wherein the ratio of said affinity-tag-binding molecules to said affinity tags present in the antisense rRNA molecules used in step b) is at least 8 to 1, such that at least a portion of said double-stranded rRNA hybrids bind to said binding matrix and are removed from said treated sample, thereby generating an rRNA-depleted sample, wherein said rRNA-depleted sample comprises at least a portion of said non-rRNA RNA molecules present in said initial sample and is substantially depleted or free of rRNA molecules that exhibit sequences exhibited by said at least one rRNA molecule.

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