http://rdf.ncbi.nlm.nih.gov/pubchem/patent/EP-1929012-A2
Outgoing Links
Predicate | Object |
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assignee | http://rdf.ncbi.nlm.nih.gov/pubchem/patentassignee/MD5_f539b0fb0491cb3849b91adfdfddd1ed |
classificationCPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N15-102 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N15-10 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N15-1031 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N15-64 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N15-66 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12N9-1252 http://rdf.ncbi.nlm.nih.gov/pubchem/patentcpc/C12P19-34 |
classificationIPCInventive | http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12N15-09 http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/C12N15-00 |
filingDate | 2006-08-11-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
inventor | http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_b339d4d6d24b40d1b4dc25e69a3e7634 http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_939f3110a2967fc28dc7f94234ac0aba |
publicationDate | 2008-06-11-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
publicationNumber | EP-1929012-A2 |
titleOfInvention | Method for in vitro recombination |
abstract | The present invention relates, e.g., to an in vitro method, using isolated protein reagents, for joining two double-stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising (a) chewing back the DNA molecules with an enzyme having an exonuclease activity, to yield single-stranded overhanging portions of each DNA molecule which contain a sufficient length of the region of sequence identity to hybridize specifically to each other; (b) specifically annealing the single-stranded overhangs; and (c) repairing single-stranded gaps in the annealed DNA molecules and sealing the nicks thus formed (ligating the nicked DNA molecules). The region of sequence identity generally comprises at least 20 non-palindromic nucleotides (nt), e.g., at least about 40 non-palindromic nt. In some embodiments of the invention, about 5% PEG is present during all steps of the reaction, and/or the repair reaction is achieved with Taq DNA polymerase and a compatible ligase, such as Taq DNA ligase. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes. It can be used, e.g., to join synthetically produced sub-fragments of a gene or genome of interest. |
isCitedBy | http://rdf.ncbi.nlm.nih.gov/pubchem/patent/US-9447445-B2 http://rdf.ncbi.nlm.nih.gov/pubchem/patent/WO-2021064106-A1 http://rdf.ncbi.nlm.nih.gov/pubchem/patent/WO-2021064115-A1 http://rdf.ncbi.nlm.nih.gov/pubchem/patent/US-9963687-B2 |
priorityDate | 2005-08-11-04:00^^<http://www.w3.org/2001/XMLSchema#date> |
type | http://data.epo.org/linked-data/def/patent/Publication |
Incoming Links
Total number of triples: 192.