Invitrogen™ MultiShot™ Stbl3™ Chemically Competent E. coli Cells, 96-well plate

Features of MultiShot Stbl3 competent cells include:

• Expected efficiency of >1 x 10⁸ transformants/μg plasmid DNA
• Optimal performance for the propagation of lentiviral vectors and maintenance of direct repeats
• Enhanced genomic DNA cloning capacity (mcrB, mrr, and hsdS20)
• Capable of producing high DNA yields

The Stbl3 strain has been essential for many applications, including genome editing CRISPR workflows, vaccine development (including SARS-CoV-2 vaccine research), cloning, and maintaining vectors with multiple repeats (e.g., gene coding protein with repeated sequence). The transformation efficiency of MultiShot Stbl3 chemically competent cells is >1 x 10⁸ cfu/μg plasmid DNA.

The Stbl3 strain is derived from the HB101 E. coli strain with a hybrid K-12 and B strain genome. It contains the recA13 allele of the recA mutation that reduces nonspecific recombination occurrence in cloned DNA. Mutations in the Stbl3 methylation-dependent restriction system (mcrB, mrr, and hsdS20) allow construction of cloning genomic DNA. This strain has endA1+, a nonspecific endonuclease I that digests DNA. The Stbl3 strain also reproducibly yields more plasmid DNA than DH strains (10-fold or more plasmid DNA than the TOP10 strain) and sustainably produces high titer lentivirus.

MultiShot Stbl3 competent cells offer:

• Expected efficiency of >1 x 10⁸ transformants/μg plasmid DNA
• Optimal performance for the propagation of lentiviral vectors and maintenance of direct repeats
• Enhanced genomic DNA cloning capacity (mcrB, mrr, and hsdS20)
• Capable of producing high DNA yields

Note: For optimal performance and to maintain plasmid with complex repeat sequences, Stbl3 cells should be cultured at 30°C. When performing plasmid DNA isolation from Stbl3 cells that have wild type endonuclease I (endA1+), ensure that buffers contain 10 mM EDTA. EDTA will inactivate the endonuclease and avoid DNA nicking and vector degradation (follow plasmid DNA purification kit instructions for endA1+ E. coli strains).

Genotype

F^–mcrB mrr hsdS20(r_B–, m_B–) recA13 supE44 ara-14 galK2 lacY1 proA2 rpsL20(StrR) xyl-5 λ–leu mtl-1