Login| Sign Up| Help| Contact|

Patent Searching and Data


Title:
ELECTROPHORETIC NUCLEIC ACID PURIFICATION METHOD
Document Type and Number:
WIPO Patent Application WO/1999/038874
Kind Code:
A2
Abstract:
An electrophoretic method for purifying a nucleic acid sample is disclosed. The method generally comprises the steps of (1) providing a nucleic acid sample comprising a desired nucleic acid and one more contaminants, (2) providing an electrophoresis matrix having a loading well and a recovery well formed therein, (3) placing the nucleic acid sample into the loading well, (4) performing a first electrophoresis comprising electrophoresing the nucleic acid sample for a first time effective to transport the desired nucleic acid out of the loading well and into the electrophoresis matrix; and (5) performing a second electrophoresis comprising electrophoresing the nucleic acid sample for a second time effective to transport the desired nucleic acid out of the electrophoresis matrix and into the recovery well. According to the method, the first and second electrophoresis steps are effective to substantially reduce the concentration of contaminants relative to the concentration of desired nucleic acid in the nucleic acid sample, thereby producing a purified nucleic acid. In the method, the loading and recovery wells may be the same or different, and the electric fields may be DC or alternating. Also disclosed is a preparative electrophoresis method employing an alternating electrical field.

More Like This:
JPS6270748METHOD FOR SEPARATING CHARGED SUBSTANCE
JP2554944[Title of Invention] Automatic Capillary Electrophoresis Device
JP2023118290PARTICLE RECOVERY METHOD AND PARTICLE RECOVERY DEVICE
Inventors:
SLATER GARY
EFCAVITCH J WILLIAM
DROUIN GUY
MAYER PASCAL
ROUSSEAU JEAN
ZHOU HONG YAN
CHIESA CLAUDIA
RUHFEL ROBERT
O'NEILL ROGER
Application Number:
PCT/US1999/001137
Publication Date:
August 05, 1999
Filing Date:
January 19, 1999
Export Citation:
Click for automatic bibliography generation   Help
Assignee:
PERKIN ELMER CORP (US)
International Classes:
B01D57/02; B03C5/00; C07H21/02; C07H21/04; C12N15/09; C12N15/10; G01N27/447; (IPC1-7): C07H1/00
Domestic Patent References:
WO1995014921A11995-06-01
WO1995014922A11995-06-01
WO1986000989A11986-02-13
Foreign References:
EP0773225A21997-05-14
EP0545689A11993-06-09
EP0644420A21995-03-22
Other References:
See also references of EP 1056758A2
Attorney, Agent or Firm:
Grossman, Paul D. (CA, US)
Vossius, Volker (Holbeinstr. 5 München, DE)
Download PDF:
View/Download PDF      PDF Help
Claims:
WE CLAIM:
1. A method for purifying a nucleic acid sample comprising the steps of : providing a nucleic acid sample comprising a desired nucleic acid and one or more contaminants; providing an electrophoresis matrix having a loading well and a recovery well formed therein; placing the nucleic acid sample into the loading well; performing a first electrophoresis comprising electrophoresing the nucleic acid sample for a first time effective to transport the desired nucleic acid out of the loading well and into the electrophoresis matrix; and performing a second electrophoresis comprising electrophoresing the nucleic acid sample for a second time effective to transport the desired nucleic acid out of the electrophoresis matrix and into the recovery well; wherein the first and second electrophoresis is effective to substantially reduce the concentration of contaminants relative the concentration of desired nucleic acid, thereby producing a purifie nucleic acid.
2. The method of claim 1 wherein the loading well and the recovery well are spatially overlapping wells.
3. The method of claim 1 wherein the loading well and the recovery well are spatially distinct wells.
4. The method of claim 2 wherein the matrix comprises a bulk portion and a wellmatrixinterface portion, the matrix being effective to trap the desired nucleic acid in the wellmatrixinterface portion during the first electrophoresis such that the desired nucleic acid is substantially prevented from entering the bulk portion of the matrix.
5. The method of claim 4 wherein the first time is less than the second time.
6. The method of claim 5 wherein the first time is equal to or greater than 20 minutes and the second time is less than or equal to 30 seconds.
7. The method of claim 4 wherein the second electrophoresis employs a LITAC electrical field.
8. The method of claim 2 wherein the first electrophoresis is sufficient to transport a portion of the contaminants out of the loading well, through the electrophoresis matrix, and into a contaminant dilution reservoir, the reservoir containing a volume of buffer sufficient to substantially dilute the contaminants entering the reservoir.
9. The method of claim 2 wherein the first electrophoresis employs a DC electrical field and the second electrophoresis employs a LITAC electrical field.
10. The method of claim 9 wherein the LITAC electrical field comprises a forward electrical field having a magnitude Ef, and a reverse electrical field having a magnitude Er7 wherein the ratio E, VEr is greater than 2.
11. The method of claim 10 wherein the LITAC electrical field comprises a forward electrical field having a magnitude Ef, and a reverse electrical field having a magnitude Er7 wherein the ratio Es/Er is approximately 2.4.
12. The method of claim 3 wherein the first electrophoresis employs a DC electrical field and the second electrophoresis employs a LITAC electrical field.
13. The method of claim 12 wherein the LITAC electrical field comprises a forward electrical field having a magnitude Ef, and a reverse electrical field having a magnitude Er7 wherein the ratio Ef/Er is greater than 2.
14. The method of claim 13 wherein the LITAC electrical field comprises a forward electrical field having a magnitude Ef, and a reverse electrical field having a magnitude Er7 wherein the ratio EdEr is approximately 2.4.
15. The method of claim 3 wherein the first electrophoresis comprises electrophoresing the desired nucleic acid in a first direction and the second electrophoresis comprises electrophoresing the desired nucleic acid in a second direction different from the first direction.
16. The method of claim 15 wherein the first electrophoresis employs a DC electrical field and the second electrophoresis employs a LITAC electrical field.
17. The method of claim 16 wherein the LITAC electrical field comprises a forward electrical field having a magnitude Ef, and a reverse electrical field having a magnitude Er7 wherein the ratio Ef/Er is greater than 2.
18. The method of claim 17 wherein the LITAC electrical field comprises a forward electrical field having a magnitude Ef, and a reverse electrical field having<BR> a magnitude Er, wherein the ratio E, VEr is approximately 2.4.
19. A method for purifying a nucleic acid sample comprising the steps of providing a nucleic acid sample comprising a desired nucleic acid and one or more contaminants; providing an electrophoresis matrix having a loading/recovery well formed therein ; wherein the electrophoresis matrix comprises a bulk portion and a well matrixinterface portion, the matrix being effective to trap the desired nucleic acid in the wellmatrixinterface portion such that the desired nucleic acid is prevented from entering the bulk portion of the matrix; placing the nucleic acid sample into the loading/recovery well; performing a first electrophoresis comprising electrophoresing the nucleic acid sample for a first time effective to transport the desired nucleic acid out of the loading/recovery well and into the wellmatrixinterface portion of the electrophoresis matrix; wherein the first electrophoresis is effective to substantially reduce the concentration of contaminants relative the concentration of desired nucleic acid, thereby producing a purifie nucleic acid; and removing the purifie nucleic acid from the loading/recovery well.
20. A method for the electrophoresis of a nucleic acid sample located in an electrophoresis matrix comprising subjecting the nucleic acid sample to a LITAC electrical field comprising a forward electrical field EF and a reverse electrical field ER.
21. The method of claim 20 wherein the ratio EF/ER ranges from 2 to 3.
22. The method of claim 21 wherein EF/ER is about 2.4.
23. A method for the electrophoresis of a nucleic acid sample located in an electrophoresis matrix comprising subjecting the nucleic acid sample to a ZIVE electrical field comprising a forward electrical field EF and a reverse electrical field ER.
24. The method of claim 23 wherein the ratio EF/ER ranges from 2 to 3.
25. The method of claim 24 wherein EF/ER is about 2.4.
Description:
INTERNATIONAL SEARCH REPORT tntet anal Application No PCT/US 99/01137 A. CLASSIFICATION OF SUBJECT MATTER IPC 6 C07H1/08 GO1N27/447 According to Intemational Patent Classification (IPC) or to both national classification and IPC B. FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) IPC 6 C07H G01N Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched Electronic data base consulted during the international search (name of data base and, where practical, search terms used) C. DOCUMENTS CONSIDERED TO BE RELEVANT Category citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. A WO 95 14921 A (PERKIN ELMER CORP; JOHNSON 1,19 BEN F (US); MENCHEN STEVEN M (US); BLOC) 1 June 1995 (1995-06-01) claims1,19 A WO 95 14922 A (PERKIN ELMER CORP) 1,19 1 June 1995 (1995-06-01) claims 1-40 A WO 86 00989 A (SCHUMACHER JUERGEN; RIESNER 1,19 DETLEV) 13 February 1986 (1986-02-13) abstract A EP 0 773 225 A (PERKIN ELMER CORP) 1,19 14 May 1997 (1997-05-14) claims 6-13 _/-- ) Further documents are listed in the continuation of box n Patent family members are listed in annex. LU Special categories of cfted documents : ° Specia) categories of cited documents : "T"later document published after the international filing date consideredto be of particular relevance invention invention "E"earlier document but published on or after the international"X"document of particular relevance; the claimed invention filing date cannot be considered novel or cannot be considered to "L"document which may throw doubts on priority claim (s) or involve an inventive step when the document is taken alone whichis cited to establish the publication date of another ^Y^ document of particular relevance ; the claimed invention citation or other special reason (as specified) cannot be considered to involve an inventive step when the "0"document referring to an oral disclosure, use, exhibition or document is combined with one or more other such docu- other means ments, such combination being obvious to a person skilled "P"document published prior to the international filing date but in the art. later than the priority date claimed"&"document member of the same patent family Date of the actual completion of the international search Date of mailing of the international search report 27 July 1999 05/08/1999 Name and mailing address of the ISA Authorized officer European Patent Office, P. B. 5818 Patentlaan 2 NL-2280 HV Rijswijk Tel. (+31-70) 340-2040, Tx. 31 651 epo nl, Fax: (+31-70) 340-3016 scout, J INTERNATIONAL SEARCH REPORT Inte onal Applicatfon No PCT/US 99/01137 C. (Continuation) DOCUMENTS CONSIDERED TO BE RELEVANT Category ° Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. A EP 0 545 689 A (UNIV NORTH CAROLINA) 1,19 9 June 1993 (1993-06-09) claims 1-11 A EP 0 644 420 A (UNIV NORTH CAROLINA; UNIV 1,19 WAKE FOREST (US)) 22 March 1995 (1995-03-22) claims 1-27 INTERNATIONAL SEARCH REPORT inte onal Applicatlon No ..,dormatlon on patent family members PCT/US 99/01137 Patent document Publication Patent cited in search report date member (s) date WO 9514921 A 01-06-1995 AU 671569 B 29-08-1996 AU 1294695 A 13-06-1995 AU 8082794 A 13-06-1995 CA 2153175 A 01-06-1995 EP 0680605 A 08-11-1995 JP 2712058 B 10-02-1998 JP 8506424 T 09-07-1996 WO 9514922 A 01-06-1995 US 5891313 A 06-04-1999 WO 9514922 A 01-06-1995 AU 671569 B 29-08-1996 AU 1294695 A 13-06-1995 AU 8082794 A 13-06-1995 CA 2153175 A 01-06-1995 EP 0680605 A 08-11-1995 JP 2712058 B 10-02-1998 JP 8506424 T 09-07-1996 WO 9514921 A 01-06-1995 US 5891313 A 06-04-1999 WO 8600989 A 13-02-1986 DE 3428385 A 20-02-1986 AT 42000 T 15-04-1989 AU 4679885 A 25-02-1986 DD 236398 A 04-06-1986 EP 0190267 A 13-08-1986 EP 0773225 A 14-05-1997 US 5891313 A 06-04-1999 AU 7063196 A 19-06-1997 CA 2189381 A 09-05-1997 JP 9201200 A 05-08-1997 EP 0545689 A 09-06-1993 US 5178737 A 12-01-1993 CA 2082906 A, C 05-06-1993 JP 2098171 C 02-10-1996 JP 6011483 A 21-01-1994 JP 8016671 B 21-02-1996 EP 0644420 A 22-03-1995 US 5453162 A 26-09-1995 CA 2131597 A 10-03-1995 JP 7167837 A 04-07-1995



 
Previous Patent: COMPOUNDS CONTAINING BORON FOR ELECTRON MICROSCOPY AND FOR BORON NEUTRON-CAPTURE THERAPY

Next Patent: PROCESS FOR SYNTHESIS OF POLYOL FATTY ACID POLYESTERS