Title:
METHOD FOR INDIRECTLY EXTRACTING NON-CULTIVABLE DNA ORGANISMS AND DNA OBTAINABLE BY SAID METHOD
Document Type and Number:
WIPO Patent Application WO2001081357
Kind Code:
A3
Abstract:
The invention concerns a method for indirectly extracting DNA target with a size greater than 300 kB from non-cultivable organisms contained in an environmental sample which consists in: grinding said sample in liquid medium; recuperating the resulting ground material; separating the organisms from the rest of the ground material by sedimentation on a cushion with density higher than 1; recuperating the organisms isolated at the surface of the cushion and incorporating it in an agarose block; extracting the DNA by lysis in the agarose block; placing the block in an agarose gel which is subjected to at least pulsed field electrophoresis; finally, recuperating the extracted DNA strands.
Inventors:
NALIN RENAUD (FR)
ROBE PATRICK (FR)
TRAN VAN VAN (FR)
ROBE PATRICK (FR)
TRAN VAN VAN (FR)
Application Number:
PCT/FR2001/000992
Publication Date:
March 14, 2002
Filing Date:
April 03, 2001
Export Citation:
Assignee:
LIBRAGEN (FR)
NALIN RENAUD (FR)
ROBE PATRICK (FR)
TRAN VAN VAN (FR)
NALIN RENAUD (FR)
ROBE PATRICK (FR)
TRAN VAN VAN (FR)
International Classes:
C12N15/10; G01N27/447; (IPC1-7): C12N15/10; C07H21/04; G01N27/447
Domestic Patent References:
WO1999008532A1 | 1999-02-25 | |||
WO1998017685A1 | 1998-04-30 | |||
WO1994022904A1 | 1994-10-13 |
Foreign References:
EP0388053A2 | 1990-09-19 | |||
EP0512181A1 | 1992-11-11 |
Other References:
AMARAL JOHN A ET AL: "Atmospheric methane consumption by forest soils and extracted bacteria at different pH values.", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 64, no. 7, July 1998 (1998-07-01), pages 2397 - 2402, XP002156716, ISSN: 0099-2240
ROEMLING U ET AL: "THE IMPACT OF TWO-DIMENSIONAL PULSED-FIELD GEL ELECTROPHORESIS TECHNIQUES FOR THE CONSISTENT AND COMPLETE MAPPING OF BACTERIAL GENOMES REFINED PHYSICAL MAP OF PSEUDOMONAS-AERUGINOSA PAO", NUCLEIC ACIDS RESEARCH, vol. 19, no. 12, 1991, pages 3199 - 3206, XP002180817, ISSN: 0305-1048
SUAU ANTONIA ET AL: "Direct analysis of genes encoding 16S rRNA from complex communities reveals many novel molecular species within the human gut.", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 65, no. 11, November 1999 (1999-11-01), pages 4799 - 4807, XP002156717, ISSN: 0099-2240
ROCHELLE P A ET AL: "A SIMPLE TECHNIQUE FOR ELECTROELUTION OF DNA FROM ENVIRONMENTAL SAMPLES", BIOTECHNIQUES, vol. 11, no. 6, 1991, pages 724, 726 - 728, XP000971960, ISSN: 0736-6205
GRIESS GARY A ET AL: "Unlimited increase in the resolution of DNA ladders.", ELECTROPHORESIS, vol. 21, no. 5, March 2000 (2000-03-01), pages 859 - 864, XP001030781, ISSN: 0173-0835
ROEMLING U ET AL: "THE IMPACT OF TWO-DIMENSIONAL PULSED-FIELD GEL ELECTROPHORESIS TECHNIQUES FOR THE CONSISTENT AND COMPLETE MAPPING OF BACTERIAL GENOMES REFINED PHYSICAL MAP OF PSEUDOMONAS-AERUGINOSA PAO", NUCLEIC ACIDS RESEARCH, vol. 19, no. 12, 1991, pages 3199 - 3206, XP002180817, ISSN: 0305-1048
SUAU ANTONIA ET AL: "Direct analysis of genes encoding 16S rRNA from complex communities reveals many novel molecular species within the human gut.", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 65, no. 11, November 1999 (1999-11-01), pages 4799 - 4807, XP002156717, ISSN: 0099-2240
ROCHELLE P A ET AL: "A SIMPLE TECHNIQUE FOR ELECTROELUTION OF DNA FROM ENVIRONMENTAL SAMPLES", BIOTECHNIQUES, vol. 11, no. 6, 1991, pages 724, 726 - 728, XP000971960, ISSN: 0736-6205
GRIESS GARY A ET AL: "Unlimited increase in the resolution of DNA ladders.", ELECTROPHORESIS, vol. 21, no. 5, March 2000 (2000-03-01), pages 859 - 864, XP001030781, ISSN: 0173-0835
Download PDF: