EP 1432797 A2 20040630 - METHODS FOR DNA MANIPULATION
Title (en)
METHODS FOR DNA MANIPULATION
Title (de)
METHODEN ZUR DNA-MUTAGENESE UND DNA-KLONIERUNG
Title (fr)
PROCEDES DE MANIPULATION D'ADN
Publication
Application
Priority
- DE 10148607 A 20011002
- EP 0211086 W 20021002
Abstract (en)
[origin: DE10148607A1] Mutagenesis and cloning of nucleic acids (NA), is new. Mutagenesis and cloning of a nucleic acid (NA), comprises: (a) preparing a circular, double-stranded (ds) NA matrix to be mutated; (b) preparing two oligonucleotides (ON) that each hybridize to one strand of the matrix and have at the 5'-ends a 2-4 base pair (bp) segment, mutually complementary, where at least one ON contains a mutated target sequence; (c) amplifying the matrix using an ON with the mutated target sequence as a primer to create a mutated ds amplicon; (d) trimming the ends of the amplicon to create a product with mutually complementary 5'-overhangs; and (e) ligating the trimmed amplicon to produce a mutated circular NA construct. Independent claims are also included for: (1) a reagent kit for the new method, comprising agents for: (i) NA amplification; (ii) trimming ds NA to create complementary ends; (iii) for ligation; and (iv) optionally for selective removal of methylated DNA; (2) cloning NA fragments into a vector; and (3) reagent kits for the method of (2).
IPC 1-7
IPC 8 full level
C12N 15/10 (2006.01); C12N 15/66 (2006.01)
CPC (source: EP US)
C12N 15/102 (2013.01 - EP US); C12N 15/66 (2013.01 - EP US)
Citation (search report)
See references of WO 03031616A2
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)
DE 10148607 A1 20030410; AU 2002342790 A1 20030422; EP 1432797 A2 20040630; US 2004248131 A1 20041209; WO 03031616 A2 20030417; WO 03031616 A3 20031211
DOCDB simple family (application)
DE 10148607 A 20011002; AU 2002342790 A 20021002; EP 0211086 W 20021002; EP 02779456 A 20021002; US 49162004 A 20040402