Talk:MiPNet14.13 Medium-MiR06: Difference between revisions
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== Preparation of MiR05 (MiR06) stock solution == | |||
* This is the version up to 2013-09-07: | |||
::1) weigh given amounts of the [[#Chemicals for MiR05|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 H<sub>2</sub>O and dissolve on a magnetic stirrer at ~30 ยฐC | |||
::4) add 120 ml of [[#Preparation of K-lactobionate stock solution|K-lactobionate stock solution]] | |||
::5) adjust the pH to 7.1 with 5 N KOH at 30 ยฐC | |||
::6) transfer the MiR05 stock solution to a 1000 ml volumentric glass flask and add H20 to a final volume of 1000 ml <span style="color:#43CD80"> = '''MiR05''' </span> | |||
::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 <span style="color:#2E8B57"> '''MiR06'''</span>, 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 ยฐC in plastic vials | |||
::<span style="color:#2E8B57"> '''MiR06''' </span> can also be prepared by adding 5 ยตl of the [[#Preparation of catalase stock solution|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. | |||
== MiPNet14.13 Medium-MiR06 page accessed == | == MiPNet14.13 Medium-MiR06 page accessed == | ||
[[MiPNet14.13 Medium-MiR06]] has been accessed more than ย | [[MiPNet14.13 Medium-MiR06]] has been accessed more than ย | ||
:*ย 5,000 times (2013-09-07) | :*ย 5,000 times (2013-09-07) |
Revision as of 09:01, 7 September 2013
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 ยฐC
- 4) add 120 ml of K-lactobionate stock solution
- 5) adjust the pH to 7.1 with 5 N KOH at 30 ยฐC
- 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 ยฐC in plastic vials
- MiR06 can also be prepared by adding 5 ยตl 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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MiPNet14.13 Medium-MiR06 has been accessed more than
- 5,000 times (2013-09-07)