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Max efficiency stbl2 cells

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Max Efficiency Stbl2 cells are a bacterial cell line designed for efficient transformation and high-yield plasmid DNA production. They are engineered to provide robust and consistent performance in cloning and plasmid amplification applications.

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Lab products found in correlation

Phusion high fidelity polymerase, by Thermo Fisher Scientific (1 mentions) Expi293 expression medium, by Thermo Fisher Scientific (1 mentions) Qubit assay kit, by Thermo Fisher Scientific (1 mentions) T7 ribomax express large scale rna production system, by Promega (1 mentions) Rneasy mini kit, by Qiagen (1 mentions)

Close spelling variants of this product

Max Efficiency Stbl2 Competent cells MAX Efficiency® Stbl2 Stbl2 cells Stbl2

3 protocols using max efficiency stbl2 cells

1

SIVmac239 Genome Optimization via Site-Directed Mutagenesis

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Primers were designed to introduce the desired point mutations into the genome using site-directed mutagenesis utilizing Phusion high fidelity polymerase (Thermo Scientific). The GeneArt Seamless PLUS kit was then used to assemble the PCR fragments of the SIVmac239 genome bearing the newly modified nucleotides (Life Technologies). The resulting plasmid was transformed into Max Efficiency Stbl2 cells (Life Technologies), expanded and purified by double banded cesium chloride centrifugation. Full-length genomic sequencing was performed on the final plasmid preps to confirm point mutation generation and correct assembly of PCR fragments. The resulting sequence-confirmed plasmid was designated SIVmac239Opt.
Fennessey C.M., Reid C., Lipkey L., Newman L., Oswald K., Piatak M J.r., Roser J.D., Chertova E., Smedley J., Gregory Alvord W., Del Prete G.Q., Estes J.D., Lifson J.D, & Keele B.F. (2015). Generation and characterization of a SIVmac239 clone corrected at four suboptimal nucleotides. Retrovirology, 12, 49.
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2

Production of Codon-Optimized PCV2 Capsid Protein

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Suspension cultures of Expi293F human cells (Life Technologies, CA) were grown in serum-free Expi293 expression medium (Life Technologies, CA) at 37°C in a 5% CO2 environment and agitated at 150 rpm in Erlenmeyer flasks. The porcine circovirus type 2 (PCV2) gene encoding the CP (GenBank: AWD32058.1) was chemically synthesized using a codon-optimized sequence by Blue Heron Technologies (Bothell, WA). The recognition site for NheI and the Kozak sequence were added right upstream from the start codon, and the recognition site for NotI was incorporated after the termination codon. The synthesized Cap gene was recovered from the transport plasmid by a double digestion with NheI and NotI restriction enzymes and sub-cloned after gel purification into the mammalian expression plasmid pcDNA3.4 cut with the same enzymes. The ligated plasmid was transformed into MAX Efficiency Stbl2 Cells (Life Technologies) and a correct clone was identified via restriction enzyme analysis and verified by sequencing.
Khayat R., Wen K., Alimova A., Gavrilov B., Katz A., Galarza J.M, & Gottlieb P. (2019). Structural characterization of the PCV2d genotype at 3.3 Å resolution reveals differences to PCV2a and PCV2b genotypes, a tetranucleotide, and an N-terminus near the icosahedral 3-fold axes. Virology, 537, 186-197.
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3

Synthesis of Transmitted Founder HIV-1

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Fourteen molecular clones corresponding to seven separate transmitted founder viruses were chemically synthesized (Blue Heron Biotech) in four or five sequential fragments with a 5′ T7 promoter, a 3′ in vitro transcription runoff site, and terminal 5′ and 3′ restriction sites to facilitate cloning into the pBR322 or pCR-XL-TOPO vectors (see Fig. S6 in the supplemental material). Plasmid DNA stock was grown in MAX Efficiency Stbl2 cells (Life Technologies), and DNA was purified with the Purelink Maxiprep (Life Technologies) according to the manufacturer’s specifications. Plasmids were linearized using the restriction enzymes described in Fig. S6 (New England Biolabs) and phenol-chloroform purified. vRNA was transcribed using the T7 RiboMAX Express large-scale RNA production system (Promega) and purified using the RNeasy minikit (Qiagen). RNA quality was assessed by agarose gel electrophoresis and quantified with the Qubit assay kit (Life Technologies).
Stoddard M.B., Li H., Wang S., Saeed M., Andrus L., Ding W., Jiang X., Learn G.H., von Schaewen M., Wen J., Goepfert P.A., Hahn B.H., Ploss A., Rice C.M, & Shaw G.M. (2015). Identification, Molecular Cloning, and Analysis of Full-Length Hepatitis C Virus Transmitted/Founder Genotypes 1, 3, and 4. mBio, 6(2), e02518-14.
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