EP 1419275 A4 20070530 - MAGNESIUM PRECIPITATE HOT START METHOD FOR MOLECULAR MANIPULATION OF NUCLEIC ACIDS
Title (en)
MAGNESIUM PRECIPITATE HOT START METHOD FOR MOLECULAR MANIPULATION OF NUCLEIC ACIDS
Title (de)
HOT START-MAGNESIUMPRÄZIPITATIONSVERFAHREN ZUR MOLEKULAREN MANIPULATION VON NUKLEINSÄUREN
Title (fr)
PROCEDE DE DEMARRAGE A CHAUD UTILISANT UN PRECIPITE DE MAGNESIUM POUR MANIPULATION MOLECULAIRE D'ACIDES NUCLEIQUES
Publication
Application
Priority
- US 0224533 W 20020802
- US 30964601 P 20010802
- US 92087201 A 20010802
- US 9178402 A 20020306
Abstract (en)
[origin: WO03012066A2] The present invention provides methods of performing enzymatic reactions, including PCR, which require the use of magnesium dependent enzymes, including restriction endonucleases, ligases, and reverse transcriptases. The method is based on sequestration of magnesium ions in the form of a precipitate which renders a magnesium dependent enzyme inactive until the appropriate time in the reaction when a certain temperature is reached and the magnesium ions are released from the precipitate. Also provided are kits comprising reagents and instructions for amplifying a target nucleic acid, for DNA digestion and ligation, and for reverse transcription ofRNA into cDNA. Furthermore, the kits and reagents of the present invention can be utilized in other reactions requiring magnesium dependent enzymes.
IPC 1-7
IPC 8 full level
C12N 15/10 (2006.01); C12P 19/34 (2006.01); C12Q 1/68 (2006.01)
CPC (source: EP)
C12N 15/10 (2013.01); C12N 15/1096 (2013.01); C12P 19/34 (2013.01); C12Q 1/6848 (2013.01); C12Q 1/686 (2013.01)
Citation (search report)
- [A] US 5250429 A 19931005 - JOLLY JAMES F [US], et al
- [A] KELLOGG ET AL: "TaqStart Antibody", BIOTECHNIQUES, INFORMA LIFE SCIENCES PUBLISHING, WESTBOROUGH, MA, US, vol. 16, no. 6, 1994, pages 1134 - 1137, XP002103159, ISSN: 0736-6205
- [PA] BARNES W M ET AL: "Magnesium precipitate hot start method for PCR", MOLECULAR AND CELLULAR PROBES, ACADEMIC PRESS, LONDON, GB, vol. 16, no. 3, June 2002 (2002-06-01), pages 167 - 171, XP004470993, ISSN: 0890-8508
- See references of WO 03012066A2
Designated contracting state (EPC)
AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LI LU MC NL PT SE SK TR
DOCDB simple family (publication)
WO 03012066 A2 20030213; WO 03012066 A3 20031204; AU 2002327421 A1 20030217; EP 1419275 A2 20040519; EP 1419275 A4 20070530
DOCDB simple family (application)
US 0224533 W 20020802; AU 2002327421 A 20020802; EP 02763410 A 20020802