http://rdf.ncbi.nlm.nih.gov/pubchem/patent/RU-2511424-C2

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assignee http://rdf.ncbi.nlm.nih.gov/pubchem/patentassignee/MD5_23022bd76b3f475342ce6ca765eab3b0
classificationIPCInventive http://rdf.ncbi.nlm.nih.gov/pubchem/patentipc/G06F19-00
filingDate 2012-07-25-04:00^^<http://www.w3.org/2001/XMLSchema#date>
grantDate 2014-04-10-04:00^^<http://www.w3.org/2001/XMLSchema#date>
inventor http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_fcbc62ea5e48cac5017980bfda88acd3
http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_4ce8406ba24261bfcf5a0a3c12a5fd48
http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_8e7bb411ee77b1e5347a385d86449a99
http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_d5798fa5bc1971d6431672be94fbe0ea
http://rdf.ncbi.nlm.nih.gov/pubchem/patentinventor/MD5_f5c5438d5c1cf1ac30da8b5b958bd1f3
publicationDate 2014-04-10-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber RU-2511424-C2
titleOfInvention Method of obtaining libraries of serial bilateral deletions by means of pcr with degenerate primer
abstract FIELD: chemistry. n SUBSTANCE: claimed invention relates to field of biotechnology. Claimed is method of constructing libraries of gene deletion derivatives based on PCR with random primer. Single-strand breaks are introduced into investigated gene in form of linear DNA by treatment with pancreatic DNAse I in a series of dilutions. Breaks are extended by treatment with polymerase I from E.coli in absence of nucleotidetriphosphates. After that, random primer, which has on 3'-tail 6-, 11- or 17-membered random sequence, and on 5'-tail - constant site (20 nucleotides), intended for adapter primer annealing is attached to matrix. After that, performed is preparative elaboration of library by PCR method with symmetric adapter primer, which is attached to prepared matrix by treatment with T4 DNA-ligase, which makes it possible to delete dimers of primers, formed due to mutual annealing of random sequences, in efficient way. Banks of deletion derivatives obtained in vivo can be further subjected to screening to select variants for their further expression in vivo. Method can be used for obtaining libraries of gene deletion derivatives for their further application in the field of biotechnology, agriculture and food, pharmaceutical industries and medicine. n EFFECT: method makes it possible to obtain shortened variants of practically important genes with improved biotechnological characteristics. n 12 dwg, 1 ex
isCitedBy http://rdf.ncbi.nlm.nih.gov/pubchem/patent/RU-2781613-C1
priorityDate 2012-07-25-04:00^^<http://www.w3.org/2001/XMLSchema#date>
type http://data.epo.org/linked-data/def/patent/Publication

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