DH10βF'DOT BioVector NTCC中国质粒载体菌种细胞基因保藏中心

DH10βF'DOT BioVector NTCC中国质粒载体菌种细胞基因保藏中心 Pirplus competent cellsFor transforming pSM2 vectors PirPlus™chemically competent E.coli DH10βpir116 and DH10βF'DOT were developed for use in rapidly transforming shRNAmir containing pSM2 vectors. The shRNAmir plasmids use the pir -dependent R6Kγ replication origin. The PirPlus E. coli express the π protein (pir gene product), which is essential for cloning, and propagation of the shRNAmir vectors. Additionally the PirPlus strains contain the pir -116 modification allowing for high copy propagation of up to 250 copies per cell compared to wild type pir+ cells1. Benefits of PirPlus™ competent cells: Pir-116 to allow for replication of plasmids containing the R6Kγ origin of replication. aliquot of 100µl are supplied with pUC19 to allow for rapid and easy transformations without wasted reactions. PirPlus™chemically competent E.coli DH10βpir116 and DH10βF'DOT yield >1 x 107 CFU/µg pUC19. PirPlus DH10βF'DOT is restriction minus mcrA D (mrr-hsdRMS-mcrBC), for cloning of methylated DNA PirPlus DH10βF'DOT contains a tonA mutation conferring resistance to T1 and T5 phages. PirPlus DH10βF'DOT is MAGIC competent for use with mating assisted or MAGIC cloning2 Strains and genotypes: DH10βpir116: DH10 b UmcC::pir116-Frt DH10βF'DOT (donor host strain) mcrA Δ (mrr-hsdRMS-mcrBC) Φ 80lacZ Δ M15 Δ lacX74 deoR recA1 endA1 ara Δ 139 Δ (ara,leu)7697 galU galK λ - rpsL nupG λ - tonA umuC::pir116-frtF'(lac + pro +Δ oriT::Tc) Kit Components: PirPlus™chemically competent E.coli DH10βpir116 and DH10βF'DOT come complete with: Chemically Competent Cells- 6 x 100µl pUC19 Control Plasmid ( 10pg/µl )- 20µl Each lot is made using optimized procedures and is tested for efficiency to ensure yields of >1x107 transformants/µg pUC19. The cells are provided in a 100µl aliquot to allow for rapid and easy transformation of your shRNA plasmid. Six aliquots are supplied together with pUC19 control vector. Shipping Information: Pirplus competent e.coli is shipped on dry ice. Store all components at -80C References: Metcalf, W.W. et al. (1994) "Use of the rep technique for allele replacement to construct new Escherichia coli hosts for maintenance of R6K gamma origin plasmids at different copy numbers" Gene 138(1-2):1-7. Li MZ and Elledge S (2005) "MAGIC, an in vivo genetic method for the rapid construction of recombinant DNA molecules" Nat Genet. 2005 Mar;37(3):311-9. Epub 2005 Jan 30