Thermo Fisher, C737303, One Shot™ Stbl3™ Chemically Competent E. coli

One Shot™ Stbl3™ Chemically Competent E. coli One Shot Stbl3 Chemically CompetentE. coliare designed for cloning direct repeats found in lentiviral expression vectors as these cells reduce the frequency of homologous recombination of direct repeats found in lentiviral vectors (ViraPower Lentiviral Expression kits) and other retroviral vectors (instability-prone HIV-based plasmids). 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 One Shot Stbl3 chemically competent cells is >1 x 108cfu/μg plasmid DNA. The Stbl3 strain is derived from the HB101E. colistrain with a hybrid K-12 and B strain genome. It contains therecA13 allele of therecA mutation that reduces nonspecific recombination occurrence in cloned DNA. Mutations in the Stbl3 methylation-dependent restriction system (mcrB,mrr,andhsdS20) allow construction of cloning genomic DNA. This strain hasendA1+, a nonspecific endonuclease I that digests DNA. The Stbl3 strain also reproducibly yields more plasmid DNA than DH strains (10-fold or more plamsmid DNA than the TOP10 strain) and sustainably produces high titer lentivirus. One Shot Stbl3 Chemically Competent E. coli offer:
• Expected efficiency of >1 x 108transformants/μg plasmid DNA
• Optimal performance for the propagation of lentiviral vectors and maintenance of direct repeats
• Enhanced genomic DNA cloning capacity (mcrB,mrr,andhsdS20)
• Capable of producing high DNA yieldsNote: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 forendA1+E. colistrains). Easy-to-use One Shot formatStbl3 Chemically CompetentE. colicells are supplied in the convenient, single-reaction One Shot format. The single-tube, single-use format allows all steps of the transformation protocol, up to plating, to take place in the same tube, thereby helping save time and prevent contamination. GenotypeF–mcrBmrr hsdS20(rB–, mB–)recA13supE44ara-14galK2lacY1proA2rpsL20(StrR)xyl-5 λ–leu mtl-1Find the strain and format that fit your needsOtherStbl strainsare available with different genetic background. The Stbl3 strain is available inMultiShot formatfor high throughput applications. Explorebacterial growthmedia formatsAntibiotic Resistance Bacterial: (Yes) Streptomycin
Blue/White Screening: No
Cloning Methylated DNA: Yes (mcrB, mrr)
Cloning Unstable DNA: Yes (recA13)
Contains F' Episome: No
High-throughput Compatibility: Low
Improves Plasmid Quality: No
Plasmid: May be used for plasmids >20 kb
Preparing Unmethylated DNA: No
Product Line: One Shot™
Product Type: Chemically Competent Cells
Quantity: 21 x 50 μL/tube
Reduces Recombination: Yes (recA13)
Shipping Condition: Dry Ice
T1 Phage - Resistant (tonA): No
Transformation Efficiency Level: Medium Efficiency (1 x 108to 1 x 109cfu/μg)
Format: Tube
Species: E. coli (K12/B hybrid)
Unit Size: Each