IP Library Granted Patent US 7,553,622
Granted Patent B2
US 7,553,622 · App. 10/500,420 · Granted Jun 30, 2009

Process for preparing processed sample liquid solution for electrophoresis

Assignee: E.I. du Pont de Nemours and Company
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Quick Facts
Patent No.
US 7,553,622
App. No.
10/500,420
Granted
Jun 30, 2009
Kind
B2
Abstract

A process for preparing a processed sample liquid solution for gel electrophoresis, comprising (a) treating a sample comprising a cell suspension in a non-shearing manner to produce a processed sample liquid solution comprising a mixture of DNA fragments extracted from the cell suspension, wherein at least one of the DNA fragments is greater than 200 kilobase pairs and (b) transferring the processed sample liquid solution in a non-shearing manner directly to an electrophoresis medium for conducting electrophoresis.

Claims (28)

1. A process for preparing a processed sample liquid solution for molecular typing of bacteria using pulsed field gel electrophoresis, comprising the steps of:

(a) treating a sample comprising a bacterial cell suspension in a non-shearing manner to produce a processed sample liquid solution comprising a mixture of DNA fragments extracted from said cell suspension, wherein at least one of said DNA fragments is greater than 200 kilobase pairs; wherein said treating comprises subjecting said cell suspension to lysis, deproteinization and digestion; and

(b) transferring said processed sample liquid solution in a non-shearing manner directly to an electrophoresis medium for conducting pulsed field gel electrophoresis.

2. The process of claim 1 wherein said cell suspension comprises one or more cells suspended in a lysis buffer.

3. The process of claim 1 wherein said lysis and deproteinization comprises treatment with the enzyme achromopeptidase.

4. The process of claim 1 wherein said digestion comprises treatment with a restriction enzyme.

5. The process of claim 4 wherein said restriction enzyme is selected from the group consisting of XbaI, SfiI SmaI, NotI, ApaI, and AscI.

6. The process of claim 1 wherein said DNA fragments are 50 kilobase pairs to 1000 kilobase pairs.

7. The process of claim 1 wherein said step (a) is automated.

8. The process of claim 1 wherein said step (b) is automated.

9. The process of claim 1 wherein said steps (a) and (b) are automated.

10. The process of claim 1 wherein said electrophoresis medium is an electrophoresis gel.

11. The process of claim 1 wherein said electrophoresis medium is a well of an electrophoresis gel.

12. The process of claim 1 wherein said electrophoresis medium is a viscous sieving solution.

13. A process for separating a mixture of DNA fragments extracted from a bacterial cell suspension for molecular typing of bacteria using pulsed-field gel electrophoresis, comprising the steps of:

(a) treating a sample comprising a bacterial cell suspension in a non-shearing manner to produce a processed sample liquid solution comprising a mixture of DNA fragments extracted from said cell suspension, wherein at least one of said DNA fragments is greater than 200 kilobase pairs; wherein said treating comprises subjecting said cell suspension to lysis, deproteinization and digestion;

(b) transferring said processed sample liquid solution in a non-shearing manner directly to an electrophoresis medium; and

(c) separating said mixture of DNA fragments by conducting pulsed field gel electrophoresis.

14. The process of claim 13 wherein said cell suspension comprises one or more cells suspended in a lysis buffer.

15. The process of claim 13 wherein said DNA fragments are 50 kilobase pairs to 1000 kilobase pairs.

16. The process of claim 13 wherein said step (a) is automated.

17. The process of claim 13 wherein said step (b) is automated.

18. The process of claim 13 wherein said steps (a) and (b) are automated.

19. The process of claim 13 wherein said processed sample liquid solution is transferred to a well of said electrophoresis medium.

20. The process of claim 13 wherein said electrophoresis is pulsed-field capillary electrophoresis.

21. The process of claim 13 wherein said lysis and deproteinization comprises treatment with the enzyme achromopeptidase.

22. The process of claim 13 wherein said digestion comprises treatment with a restriction enzyme.

23. The process of claim 22 wherein said restriction enzyme is selected from the group consisting of XbaI, SfiI, SmaI, NotI, ApaI, and AscI.

Assignments (6)
RELEASE OF SECURITY INTEREST Recorded Mar 18, 2021
From: ARES CAPITAL CORPORATION, AS ADMINISTRATIVE AGENT
To: QUALICON DIAGNOSTICS LLC
Reel/Frame 055644/0636 →
TERMINATION AND RELEASE OF SECURITY INTEREST IN PATENTS AT REEL/FRAME NO. 41983/0340 Recorded Mar 16, 2021
From: ANTARES CAPITAL LP, AS ADMINISTRATIVE AGENT
To: QUALICON DIAGNOSTICS LLC
Reel/Frame 055628/0366 →
SECURITY INTEREST Recorded Mar 16, 2017
From: QUALICON DIAGNOSTICS LLC, AS THE GRANTOR
To: ARES CAPITAL CORPORATION, AS SECOND LIEN ADMINISTRATIVE AGENT
Reel/Frame 041602/0334 →
FIRST LIEN PATENT SECURITY AGREEMENT Recorded Mar 13, 2017
From: QUALICON DIAGNOSTICS LLC
To: ANTARES CAPITAL LP, AS FIRST LIEN ADMINISTRATIVE AGENT
Reel/Frame 041983/0340 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 5, 2017
From: E. I. DU PONT DE NEMOURS AND COMPANY
To: QUALICON DIAGNOSTICS LLC
Reel/Frame 041468/0214 →
MERGER Recorded Feb 18, 2005
From: QUALICON INC.
To: E.I. DU PONT DE NEMOURS
Reel/Frame 015687/0838 →
Continuity (1)
Related Publication 20050029104A1 · Feb 10, 2005