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TOP10 chemically competent cells: Difference between revisions
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TOP10 chemically competent cells (view source)
Revision as of 11:13, 27 November 2006
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* Transformation efficiency is 15 x colony count x 10<sup>5</sup> | * Transformation efficiency is 15 x colony count x 10<sup>5</sup> | ||
* There is anecdotal evidence that refreezing partially used cell aliquots dramatically reduces transformation efficiency. | * There is anecdotal evidence that refreezing partially used cell aliquots dramatically reduces transformation efficiency. | ||
==5x Ligation Adjustment Buffer== | |||
* Intended to be mixed with ligation reactions to adjust buffer composition to be near the CCMB80 buffer | |||
* KOAc 40 mM (40 ml/liter of 1 M KOAc solution, pH 7.0) | |||
* CaCl<sub>2</sub> 400 mM (200 ml/l of a 2 M solution) | |||
* MnCl<sub>2</sub> 100 mM (100 ml/l of a 1 M solution) | |||
* Glycerol 46.8% (468 ml/liter) | |||
* pH adjustment with 2.3% of a 10% acetic acid solution (23 ml/liter) | |||
* Test pH adjustment by mixing 4 parts ligation buffer + 1 part 5x ligation adjustment buffer and checking pH to be 6.3 - 6.5 | |||
==References== | ==References== | ||
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# Jesse05 US Patent 6,960,464 [[Media:pat6960464.pdf]] | # Jesse05 US Patent 6,960,464 [[Media:pat6960464.pdf]] | ||
</biblio> | </biblio> | ||
