Max efficiency stbl2

Manufactured by Thermo Fisher Scientific

The MAX Efficiency® Stbl2 is a laboratory product from Thermo Fisher Scientific. It is designed for bacterial transformation applications. The core function of the MAX Efficiency® Stbl2 is to facilitate the introduction of DNA into competent bacterial cells.

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E. coli MAX Efficiency Stbl2 Max Efficiency Stbl2 Competent cells Max Efficiency Stbl2 cells Stbl2

2 protocols using max efficiency stbl2

Cloning and Purification of pK201/CAT Plasmid

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The high-copy PK201/CAT plasmid, a kind gift of E. Di Mauro (University of Rome, “La Sapienza”), is 3228 base pairs long and contains the StuI–AccI 211-bp bent segment from the Trypanosomatidae protozoan C. fasciculata cloned in the BamHI site of the vector pSP65 [18 (link)].
E. coli competent cells (MAX Efficiency® Stbl2™, Invitrogen™) were transformed with plasmid DNA by means of the heat shock procedure and selected by ampicillin. Pelleted bacteria were treated with the alkaline lysis procedure to purify the pK201/CAT plasmid DNA (pDNA) [30 (link)]. The quality of the preparation was checked by 1 % (w/v) agarose gel electrophoresis, and the concentration and purity assessed by determining the A260/280 and A260/320 ratios in UV-Vis spectrometry.

Panza F., Alcaro M.C., Petrelli F., Angelotti F., Pratesi F., Rovero P, & Migliorini P. (2016). A novel DNA/histone H4 peptide complex detects autoantibodies in systemic lupus erythematosus sera. Arthritis Research & Therapy, 18, 220.

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Molecular Cloning and Yeast Culture Protocols

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E. coli DH10β was used for all molecular cloning except for plasmids containing the LacO or TetO array, which were carried with MAX Efficiency^® Stbl2™ (Invitrogen) and NEB^® Stable Competent E. coli (New England BioLabs Inc.), respectively. LB medium containing 50 μg mL⁻¹ of ampicillin was used to grow DH10β while SOC medium was used to grow Stbl2 and Stable Competent cells. DH10β cells were grown at 37 °C while Stbl2 and Stable Competent cells were grown at 30 °C.
S. cerevisiae strains used in this study are listed in Tables S1–2. All modified yeast strains were isogenic to BY4741. Yeast rich medium (YPD) and synthetic complete medium (SD) containing 0.67% yeast nitrogen base, complete supplement mixture (CSM) with appropriate dropout, and 2% glucose were used to culture yeast.

Lin J.L., Ekas H., Deaner M, & Alper H.S. (2019). CRISPR-PIN: Modifying gene position in the nucleus via dCas9-mediated tethering. Synthetic and Systems Biotechnology, 4(2), 73-78.

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