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One shot stbl3 e coli

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One Shot Stbl3 E. coli is a chemically competent bacterial strain used for transformation and cloning applications. It is designed to provide reliable and efficient transformation of DNA into E. coli cells.

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Lab products found in correlation

Plasmid maxiprep kit, by Qiagen (1 mentions) Plvx puro lentiviral plasmid, by Takara Bio (1 mentions)

2 protocols using one shot stbl3 e coli

1

Lentiviral Plasmid Construction: SV40 Large T Antigen

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The temperature-sensitive large T antigen SV40 sequence, SVU19tsa58, was digested from pZipSVU19tsa58 plasmid (a gift from Parmjit Jat, UCL, UK) using the BamH1 restriction site as previously described10 (link),11 (link). The sequence was ligated into the pLVX Puro lentiviral plasmid (Clontech, CA) using the same restriction sites and sequenced to confirm correct orientation. Plasmid DNA was amplified in One Shot Stbl3 E. coli (Thermo-Fisher Scientific) and purified using a Plasmid Maxiprep kit (Qiagen).
Prideaux M., Wright C.S., Noonan M.L., Yi X., Clinkenbeard E.L., Mevel E., Wheeler J.A., Byers S., Wijenayaka A.R., Gronthos S., Sankar U., White K.E., Atkins G.J, & Thompson W.R. (2021). Generation of two multipotent mesenchymal progenitor cell lines capable of osteogenic, mature osteocyte, adipogenic, and chondrogenic differentiation. Scientific Reports, 11, 22593.
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2

Constructing CBP Mutant Variants

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To clone full-length CBPdelta-loop and CBPmutant loop into pTRE3G, constructs were generated using the same method as for CBP-HAT domain constructs (see above) in pAcGFP-C1-CBP3465–7326. Mutant CBP3465–7326 was then sub-cloned into SalI/BamHI sites in pAcGFP-C1-CBP1–3464 to reconstruct FL-CBP in frame with the N-terminal GFP-tag. Cloning re-forms the silent SalI site in full-length CBP; all constructs were checked for correct frame by sequencing through restriction sites (plasmid: pAcGFPC1:CBP-FLwt; pAcGFPC1:CBP-FLdelta-loop; pAcGFPC1:CBP-FLmutant-loop). N-terminal GFP-tagged FL-CBP was sub-cloned with an additional in-frame N-terminal Flag-epitope tag into BamHI/NotI sites in the pTRE3G vector using primers (Fwd: DB_PCR_17; Rvs: DB_PCR_18) (plasmid: pTRE3G:CBP-FLwt; pTRE3G:CBP-FLdelta-loop; pTRE3G:CBP-FLmutant-loop). All work with viral vectors was carried out using One Shot Stbl3 E. coli (ThermoFisher) to reduce recombination of repeat sequences.
Bose D.A., Donahue G., Reinberg D., Shiekhattar R., Bonasio R, & Berger S.L. (2017). RNA binding to CBP stimulates histone acetylation and transcription. Cell, 168(1-2), 135-149.e22.
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