Treatment of paraffin embedded tissue for gene analysis

• US 5,672,696 A
• Filed: 07/05/1995
• Issued: 09/30/1997
• Est. Priority Date: 07/06/1994
• Status: Expired due to Fees

• Current Assignee: WAKO Pure Chemical Industries Limited (Fujifilm Holdings Corporation)
• Original Assignee: WAKO Pure Chemical Industries Limited (Fujifilm Holdings Corporation)
• Inventors: Wang, Lu, Hirayasu, Kazunari
• Primary Examiner(s): Kunz, Gary L.
• Application Number:
  US08/498,775
• Time in Patent Office: 818 Days
• Field of Search: 536/25.41, 536/25.42, 536/25.4
• US Class Current: 536/25.42
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2521/537 Protease
• CPC Class Codes: C12Q 2523/113 Denaturating agents
• CPC Class Codes: C12Q 2527/101 Temperature

Abstract

Rapid and easy preparation of a sample for a gene analysis or high-purity nucleic acid suitable for gene amplification especially, for example, by the PCR method is made possible by a process for treating a paraffin-embedded tissue sample to be used for a gene analysis, which comprises heating an aqueous suspension containing a surfactant having a protein-denaturational action and a deparaffinized tissue sample obtained from a paraffin-embedded tissue sample at 60° C. or higher, or the above-mentioned process which further comprises reacting the heat-treated aqueous solution with a protease, or the above-mentioned process which further comprises reacting the heat-treated aqueous solution with a protease, mixing the resulting reaction solution with a solution containing an organic compound having a protein-denaturational action other than the aforesaid surfactant, and precipitating nucleic acid from the resulting reaction solution by addition of an alcohol.

Claims

1. A process for treating a paraffin-embedded tissue sample to be used for a gene analysis, which comprises

heating an aqueous suspension containing a surfactant having a protein-denaturational action and a deparaffinized tissue sample obtained from a paraffin-embedded tissue sample at 60°

C. or higher,

reacting the heat-treated aqueous suspension with a protease,

mixing the resulting reaction solution with a solution containing hydroxybenzoic acid.

2. A process for treating a paraffin-embedded tissue sample to be used for a gene analysis, which comprises

heating an aqueous suspension containing a surfactant having a protein-denaturational action and a deparaffinized tissue sample obtained from a paraffin-embedded tissue sample at 60°

C. or higher.

3. A process according to claim 2, wherein heating is carried out at 70°

C. to a boiling point of the aqueous suspension.

4. A process for treating a paraffin-embedded tissue sample to be used for a gene analysis, which comprises

heating an aqueous suspension containing a surfactant having a protein-denaturational action and a deparaffinized tissue sample obtained from a paraffin-embedded tissue sample at 60°

C. or higher, and

reacting the heat-treated aqueous suspension with a protease.

5. A process for treating a paraffin-embedded tissue sample to be used for a gene analysis, which comprises

heating an aqueous suspension containing a surfactant having a protein-denaturational action and a deparaffinized tissue sample obtained from a paraffin-embedded tissue sample at 60°

C. or higher,

reacting the heat-treated aqueous suspension with a protease, and

precipitating a nucleic acid from the resulting reaction solution.

7. A process according to claim 5, wherein the solution containing an organic compound is an aqueous alcoholic solution containing an organic compound having one or more phenolic hydroxyl groups.

6. A process for treating a paraffin-embedded tissue sample to be used for a gene analysis, which comprises

heating an aqueous suspension containing a surfactant having a protein-denaturational action and a deparaffinized tissue sample obtained from a paraffin-embedded tissue sample at 60°

C. or higher,

reacting the heat-treated aqueous suspension with a protease,

mixing the resulting reaction solution with a solution containing an organic compound having a protein-denaturational action other than the surfactant mentioned above, and

precipitating a nucleic acid from the resulting reaction solution.

8. A kit for treating a paraffin-embedded tissue sample to be used for a gene analysis, which comprises

a container containing a surfactant having a protein-denaturational action,

a container containing a protease, and

a container containing a hydroxybenzoic acid solution.

9. A kit according to claim 8, wherein the surfactant is sodium dodecylsulfate and the protease is papain.

10. A process for treating a paraffin-embedded tissue sample to be used for a gene analysis, which comprises

heating an aqueous suspension containing a surfactant having a protein-denaturational action and a deparaffinized tissue sample obtained from a paraffin-embedded tissue sample,

reacting the heat-treated aqueous suspension with a protease,

mixing the resulting reaction solution with an aqueous alcoholic solution containing hydroxybenzoic acid, and

precipitating a nucleic acid from the resulting reaction solution.