Talk:MiPNet14.13 Medium-MiR06
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Revision as of 08:16, 30 October 2013 by Gnaiger Erich (talk | contribs)
Preparation of MiR05 (MiR06) stock solution
- This is the version up to 2013-09-07:
- 1) weigh given amounts of the listed chemicals (except BSA and lactobionic acid) and transfer to a 1000 ml glass beaker
- 2) disrupt big lumps mechanically. It is recommended to do this before adding water, because during dissolution these lumps do not disintegrate easily.
- 3) add ~800 ml H2O and dissolve on a magnetic stirrer at ~30 °C
- 4) add 120 ml of K-lactobionate stock solution
- 5) adjust the pH to 7.1 with 5 N KOH at 30 °C
- 6) transfer the MiR05 stock solution to a 1000 ml volumentric glass flask and add H20 to a final volume of 1000 ml = MiR05
- 7) check pH again and adjust if necessary with small amounts of 5 N KOH
- 8) dissolve the BSA in a subsample of the MiR05 stock solution and add to the final MiR05 (the separate preparation of the BSA solution is recommended, since BSA produces foams that do not dissolve easily)
- 9) to prepare MiR06, add 280 000 units of catalase (100 mg of catalase powder containing 2800 u/mg solid) per 1 L MiR05 (280 units / ml final concentration)
- 10) divide into 40 ml portions and store at -20 °C in plastic vials
- MiR06 can also be prepared by adding 5 µl of the catalase stock solution directly into the O2k-chamber filled with MiR05 at the start of the experiment. The final catalase concentration in the 2 ml Oxygraph-2k chamber = 280 u/ml.
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