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Calcium chloride (CaCl2) transformation is a laboratory technique in prokaryotic (bacterial) cell biology. [1] . The addition of calcium chloride to a cell suspension promotes the binding of plasmid DNA to lipopolysaccharides (LPS).
The divalent cations generate coordination complexes with the negatively charged DNA molecules and LPS. DNA, being a larger molecule, cannot itself cross the cell membrane to enter into the cytosol. The heat shock step strongly depolarizes the cell membrane of CaCl2-treated cells.
15 Ιουλ 2024 · In the intricate world of molecular biology, achieving efficient DNA transformation is crucial for a myriad of downstream applications. Enter calcium chloride dihydrate (CaCl2·2H2O), a seemingly simple compound that plays a pivotal role in this fundamental laboratory technique.
15 Μαΐ 2015 · Calcium chloride solution is added at ice-cold temperature to enhance brining DNA and LPS molecules in proximity, mediated through Ca2+, by reducing the degree of disorder in the system.
18 Νοε 2010 · These theoretical results are strongly supported by revisiting crystallographic structures that contain bpDNase I. One Ca 2+ stabilizes the functional DNase I structure. The presence of Mg 2+ in close vicinity to the catalytic pocket of bpDNase I reinforces the idea of a cation-assisted hydrolytic mechanism. Importantly, Poisson-Boltzmann-type ...
The calcium bound to the DNA further helps the DNA to adsorb to the competent cell (Panja et al., 2008b). Moreover, DNA binding proteins present in the cell membrane could also aid in this interaction. The anchorage of the DNA to the membrane eliminates the risk of detachment or expulsion of DNA (Clark et al., 2002). Further, the low ...
30 Νοε 2009 · Hanahan's method was found to be most effective for DH5α, XL-1 Blue and JM109 strains (p <0.05), whilst the CaCl2 method was best for SCS110, TOP10 and BL21 strains (p <0.05).
