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US 2019/0169598 A1Published application
Methods and compositions for isolating small rna
AssigneeApplied Biosystems, LLC
InventorRichard CONRAD
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Abstract
The present invention concerns the use of methods and compositions for the isolation of small RNA molecules (100 nucleotides or fewer), such as microRNA and siRNA molecules. Such molecules are routinely lost in commonly used isolation procedures and therefore the present invention allows for a much higher level of enrichment or isolation of these small RNA molecules.
Claims19
-53 (canceled)
A method for isolating small RNA molecules having at most 100 nucleotides or fewer from a sample comprising:
- a) lysing cells in a lysing solution to produce a lysate;
- wherein the lysing solution comprises a chaotropic agent;
- b) adding to the lysate an ethanol solution to form a lysate/ethanol mixture;
- c) applying the lysate/ethanol mixture to a first solid support and collecting flow-through lysate/ethanol mixture;
- d) adding to the flow-through lysate/ethanol mixture of step c) an ethanol solution;
- e) applying the lysate/ethanol mixture of step d) to a second solid support;
- and
- f) eluting small RNA molecules from the solid support, wherein the lysate/ethanol mixture applied to the first solid support is between about 20% to about 35% ethanol and wherein the lysate/ethanol mixture applied to the second solid support is between about 35% to about 70% ethanol.
5 more elements
Dependencies are inferred from the claim text.