Magnesium aminoclays as plasmid delivery agents for non-competent Escherichia coli JM109 transformation
نویسندگان
چکیده
Magnesium aminoclays were synthesized and used to transform non-competent Escherichia coli JM109 using the exogenous plasmid pUC19. The structure determined for Mg is analogous 2:1 trioctahedral smectites such as talc, with an approximate composition R8Si8Mg6O16(OH)4, where R = CH2CH2NH2, morphologically arranged in layered sheets. employed a cationic vehicle that enabled passage of plasmids across cell envelope led genetic modification host. A stock solution 10 mg/mL was prepared, mixed E. pUC19 plasmid, spread over Petri dishes containing lysogeny broth (LB), isopropyl β-D-1-thiogalactopyranoside (IPTG), 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal), ampicillin various concentrations agar (1–4%). transformation efficiency obtained higher 1% 2% even though also occurred at 3% 4%. optical density spreading time adjusted, favoring when cells their exponential growth phase (OD600 1.0) 90 s. Transformation confirmed by blue colonies LB/IPTG/X-gal/agar ampicillin, regrowth biomass liquid media agarose gel electrophoresis linearized followed plasmidic DNA extraction from 4 colonies. maximum achieved 7.0 × 103 CFU/μg This approach proved be suitable convenient, cost-effective, room-temperature, risk-free rapid JM109.
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ژورنال
عنوان ژورنال: Applied Clay Science
سال: 2021
ISSN: ['1872-9053', '0169-1317']
DOI: https://doi.org/10.1016/j.clay.2021.106010