TG1 Phage Competent™ Cells
Technical Description
TG1 Phage-Competent™ cells are concentrated stabilized bacterial TG1 cells. Phage-competent cells can undergo multiple freeze-thaw cycles without losing their titer and can be expanded without latency to generate large volumes of cells ready for transduction. Phage infectivity is usually equal or higher than infectivity measured on bacteria freshly prepared and transduction gives more reliable and reproducible numbers of cfu and pfu.
Applications
Phage display.
For research use only; not intended for any animal or human therapeutic or diagnostic use.
Specifications
Composition: TG1 supE thi-1 Δ(lac-proAB) Δ(mcrB-hsdSM)5, (rK-mK-), F' [traD36 proAB+ lacIq lacZΔM15] in freezing medium.
Titer: 20 OD600/ml, >2E10 cfu/ml
Doubling Time: ~30 min at 37ºC and 250 rpm in 2xYT medium
Storage temperature: -80°C.
Product size: 10x 0.5 ml.
Quality Control & Certification of Analysis
Concentration:
Absorption is measured at 600 nm in 1 cm path cuvette after dilution in freezing medium.
Titer:
Titer in cfu/ml is determined by plating appropriate dilutions in 2xYT medium on agar plates without antibiotics.
Contaminations:
No colonies are found after plating 50 µl of cells on 2xYT agar plates supplemented with either ampicillin 150 µg/ml, kanamycin 75 µg/ml, tetracycline 20 µg/ml or chloramphenicol 34 µg/ml. There is no observable growth in 2xYT liquid medium at 37ºC in the presence of kanamycin 75 µg/ml. Bacteria grew o/n in liquid medium do not show any sign of lysis and do no have any precipitable M13 bacteriophage.
Certification:
Products meet all specifications.
Notes
Thawing & Freezing Phage-Competent™ bacteria:
Always keep the cells at -80ºC in their original packaging. When needed, take out a tube, incubate at 37ºC in a water bath between 2 min and 5 min and immediately transfer on ice. When done, put the cells back in their original packaging at -80ºC. All testing were done with storage on ice no longer than 15 min and showed that cells could be thawed and frozen at least 5 times without losing their titer or their transducibility. Thawing the cells on ice will see a rapid decrease of the titer.
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Citations
Liu Z, Kim D, Kang S, Jung JU. (2024). A Detailed Protocol for Constructing a Human Single-Chain Variable Fragment (scFv) Library and Downstream Screening via Phage Display. Methods and Protocols. 2024; 7(1):13.
Jeong SL, Zhang H, Yamaki S et al. (2022). Nucleic Acids Res.,2:gkac995.
Jester BW, Zhao H, Gewe M, Adame T. et al. (2022). Development of spirulina for the manufacture and oral delivery of protein therapeutics. Nat. Biotechnol.,40(6):956-964.
Salem, R., El-Kholy, A.A., Ibrahim, M. (2019). Eight novel single chain antibody fragments recognising VP2 of foot-and-mouth disease virus serotypes A, O, and SAT 2. Virology, 533:145-154.