Method and system for RNA analysis by matched ion polynucleotide chromatography
AssigneeTransgenomic, Inc.
InventorsDouglas T. Gjerde, David P. Hornby, Christopher P. Hanna, Alexander I. Kuklin
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Abstract
A Matched Ion Polynucleotide Chromatography method and system for size-based segregation of a mixture of RNA molecules. The method includes applying the mixture to a reverse phase column containing polymeric beads and eluting, preferably under denaturing conditions, the RNA molecules with a mobile phase which includes counterion reagent and an organic component. The method can be used in segregating RNA molecules having lengths in the range of about 100 to 20,000 nucleotides. Improved segregation is obtained using a chromatography column having an ID greater than about 5 mm.
Claims32
A chromatographic method for segregating a mixture of RNA molecules having lengths exceeding about 100 nucleotides, said method comprising:
- a) applying a solution of said fragments and counterion reagent to a column containing polymeric beads having non-polar surfaces, wherein said beads have an average diameter of about 1 to about 100 microns, wherein said surfaces are substantially free from multivalent cations that are free to interfere with RNA segregation;
- b) eluting said RNA molecules with a mobile phase which includes said counterion reagent and an organic component.
An improved method for segregating a mixture of RNA molecules by Matched Ion Polynucleotide Chromatography, said mixture comprising molecules having lengths exceeding about 100 nucleotides, the method comprising:
- a) applying a solution of said molecules and counterion reagent to a column containing polymeric separation beads having non-polar surfaces, wherein said separation beads have an average diameter of 1 to 100 microns, said column having an ID greater than about 5 mm, wherein said surfaces are substantially free from multivalent cations that are free to interfere with RNA segregation;
- b) eluting said RNA molecules with a mobile phase which includes said counterion reagent and an organic component.
Dependencies are inferred from the claim text.