Methods and reagents for preserving RNA in cell and tissue samples

• US 8,178,296 B2
• Filed: 08/03/2009
• Issued: 05/15/2012
• Est. Priority Date: 07/31/1998
• Status: Expired due to Fees

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Lader, Eric S.
• Primary Examiner(s): BAUSCH, SARAE L
• Application Number:
  US12/534,182
• Publication Number:
  US 20100028852A1
• Time in Patent Office: 1,016 Days
• Field of Search: None
• US Class Current: 435/6.1
• CPC Class Codes: C12N 1/04 Preserving or maintaining viable microorganisms immobilised microorganisms C12N11/00
• CPC Class Codes: C12N 15/1003 Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence

Abstract

This specification relates to the field of molecular biology and provides novel methods and reagents for preserving and protecting the ribonucleic acid (RNA) content of samples from degradation prior to RNA isolation. This preservation may be accomplished without ultra-low temperature storage or disruption of the tissue.

Claims

1. A method of preserving RNA in an intact cell of a biological sample comprising:

preserving the RNA of an intact cell by contacting the intact cell of the sample with an RNA preservation composition comprising a chelator of divalent cations;

a citrate buffer or an acetate buffer; and

an ammonium sulfate salt at a concentration between 20 g/100 ml and a saturating concentration of the salt, wherein the intact cell remains intact and the RNA in the intact cell is preserved thereby.

2. The method of claim 1, wherein the sample is a suspension of cells.

3. The method of claim 1, wherein the sample is a solid tissue sample.

4. The method of claim 1, wherein the sample is a blood sample.

5. The method of claim 1, wherein the sample is a water sample.

6. The method of claim 1, wherein the sample comprises a pathogen within a tissue sample or other organism.

7. The method of claim 1, further comprising isolating preserved RNA.