PirPlus™chemically competent E.coli DH10βpir116  and DH10βF'DOT  were developed for use in rapidly transforming  shRNA^mir 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 shRNA^mir 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.