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TOP10 chemically competent cells: Difference between revisions
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TOP10 chemically competent cells (view source)
Revision as of 13:50, 25 January 2007
, 25 January 2007→5x Ligation Adjustment Buffer
Barry Canton (talk | contribs) No edit summary |
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* MnCl<sub>2</sub> 100 mM (100 ml/l of a 1 M solution) | * MnCl<sub>2</sub> 100 mM (100 ml/l of a 1 M solution) | ||
* Glycerol 46.8% (468 ml/liter) | * Glycerol 46.8% (468 ml/liter) | ||
* pH adjustment with 2.3% of a 10% acetic acid solution (23 ml/liter) | * pH adjustment with 2.3% of a 10% acetic acid solution (12.8ml/liter) | ||
** Previous protocol indicated amount of acetic acid added should be 23 ml/liter but that amount was found to be 2X too much per tests on 1.23.07 --[[User:Meaganl|Meaganl]] 15:50, 25 January 2007 (EST) | |||
* water to 1 liter | * water to 1 liter | ||
* autoclave or sterile filter | * autoclave or sterile filter | ||
* Test pH adjustment by mixing 4 parts ligation buffer + 1 part 5x ligation adjustment buffer and checking pH to be 6.3 - 6.5 | * 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== | ||
