EP 1086250 A2 20010328 - METHOD OF GENERATING NUCLEIC ACID HYBRIDS FOR MUTATION ANALYSIS
Title (en)
METHOD OF GENERATING NUCLEIC ACID HYBRIDS FOR MUTATION ANALYSIS
Title (de)
VERFAHREN ZUR HERSTELLUNG VON NUKLEINSÄUREHYBRIDEN ZUR MUTATIONSANALYSE
Title (fr)
Procédé pour générer des hybrides d'acides nucléiques pour l'analyse de mutations
Publication
Application
Priority
- GB 9901691 W 19990614
- GB 9812674 A 19980612
Abstract (en)
[origin: WO9964624A2] A method of producing a hybrid DNA molecule allowing the assembly of sequences x1, x2.....xn where n is greater than or equal to 3 (e.g. give sequences) from diverse locations into a hybrid molecule for the purpose of mutation analysis. The method comprising the steps of: (1) providing in a single reaction mixture: (a) the sequences x1, x2.......xn and their complementary sequences x1', x2'.........xn', to be assembled into the hybrid molecule; (b) for each pair of complementary sequences defined in (a) a respective pair of PCR primers each having a priming sequence and which are such that the primers hybridising to the 3' ends of any two sequences (xi, x'(i+1)), where i is 1 to (n-1), have specifically complementary linker sequences; (2) effecting a first stage PCR reaction in which those primers provided with linker sequences are present in limiting concentrations; and (3) effecting a second stage PCR reaction using a single pair of primers one of which provides the 5'-end of the sense strand and other of which provides the 3'-end of the anti-sense strand of the required hybrid molecule; whereby said hybrid molecule is generated.
IPC 1-7
IPC 8 full level
C12N 15/09 (2006.01); C12N 15/10 (2006.01); C12Q 1/68 (2006.01); C12Q 1/6811 (2018.01); C12Q 1/6853 (2018.01)
CPC (source: EP)
C12N 15/1093 (2013.01); C12Q 1/6811 (2013.01); C12Q 1/6853 (2013.01)
Citation (search report)
See references of WO 9964624A2
Designated contracting state (EPC)
BE CH DE DK ES FI FR GB IE IT LI NL SE
DOCDB simple family (publication)
WO 9964624 A2 19991216; WO 9964624 A3 20000914; AU 5377599 A 19991230; CA 2330252 A1 19991216; EP 1086250 A2 20010328; GB 9812674 D0 19980812; JP 2002517258 A 20020618
DOCDB simple family (application)
GB 9901691 W 19990614; AU 5377599 A 19990614; CA 2330252 A 19990614; EP 99939503 A 19990614; GB 9812674 A 19980612; JP 2000553614 A 19990614