Polynucleotide capture materials, and methods of using same

• US 10,100,302 B2
• Filed: 12/21/2015
• Issued: 10/16/2018
• Est. Priority Date: 07/13/2007
• Status: Active Grant

• Current Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Original Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Inventors: Brahmasandra, Sundaresh N., Craig, Elizabeth
• Primary Examiner(s): Crane, Lawrence E
• Application Number:
  US14/976,746
• Publication Number:
  US 20160102305A1
• Time in Patent Office: 1,030 Days
• CPC Class Codes: C07C 233/36 having the carbon atom of the carboxamide group bound to a hydrogen atom or to a carbon atom of an acyclic saturated carbon skeleton
• CPC Class Codes: C07H 1/08 from natural products
• CPC Class Codes: C08G 83/003 Dendrimers
• CPC Class Codes: C12N 15/1013 by using magnetic beads
• CPC Class Codes: C12Q 1/6813 Hybridisation assays

Abstract

Methods for processing polynucleotide-containing biological samples, and materials for capturing polynucleotide molecules such as RNA and/or DNA from such samples. The RNA and/or DNA is captured by polyamindoamine (PAMAM (Generation 0)) bound to a surface, such as the surface of magnetic particles. The methods and materials have high efficiency of binding RNA and of DNA, and of release, and thereby permit quantitative determinations.

Claims

1. A kit, comprising:

a lysis buffer;

binding particles having PAMAM(Generation

     0) molecules covalently bound thereto;

a wash solution; and

a release solution having a pH of approximately 9 or higher.

2. The kit of claim 1, wherein one amine group on the PAMAM molecules covalently bonds with the carboxyl groups on the surface of the binding particle.

3. The kit of claim 1, wherein a plurality of amine groups on the PAMAM molecules covalently bond with the carboxyl groups on the surface of the binding particle.

4. The kit of claim 1, wherein the binding particles are magnetic.

5. The kit of claim 1, wherein the PAMAM molecules are made from an ethylene diamine monomer core.

6. The kit of claim 1, wherein the PAMAM molecules on the binding particles consist of five amine groups available for protonation.

7. The kit of claim 1, wherein the release solution has a volume of less than 3 microliters.

8. The kit of claim 1, wherein the release solution comprises Tris.

9. The kit of claim 1, wherein the release solution comprises bis-tris-propane.

10. The kit of claim 1, wherein the wash solution comprises Tris-EDTA and 1% Triton X 100 at pH 8.0.

11. The kit of claim 1, wherein the lysis buffer has a pH between 4 and 8.

12. The kit of claim 1, wherein the release solution has a pH of approximately 11 or higher.

13. The kit of claim 1, wherein the release solution has a pH of approximately 12.

14. The kit of claim 1, wherein the PAMAM on the binding particles have a molecular weight in the range of 500-600 Da.

15. The kit of claim 14, wherein the PAMAM on the binding particles have a molecular weight of approximately 516 Da.

16. The kit of claim 1, wherein the PAMAM molecules are made from a monomer core selected from the group consisting of 1,2-propylene diamine, 1,3-propylene diamine, 1,2-butylene diamine, 1,3-butylene diamine, and 1,4-butylenediamine.

17. The kit of claim 1, wherein the PAMAM molecules are made from an ethylene diamine monomer core.