Difference between revisions of "Talk:MiPNet14.13 Medium-MiR06"
From Bioblast
| Line 1: | Line 1: | ||
== Preparation of MiR05 (MiR06) stock solution == | == Preparation of MiR05 (MiR06) stock solution == | ||
* This is the version up to 2013-09-07: | * This is the version up to 2013-09-07: | ||
::1) | ::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) | ::2) Disrupt big lumps mechanically. It is recommended to do this before adding water, because during dissolution these lumps do not disintegrate easily. | ||
::3) | ::3) Add ~800 ml H<sub>2</sub>O and dissolve on a magnetic stirrer at ~30 °C | ||
::4) | ::4) Add 120 ml of [[#Preparation of K-lactobionate stock solution|K-lactobionate stock solution]] | ||
::5) | ::5) Adjust the pH to 7.1 with 5 N KOH at 30 °C | ||
::6) | ::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) | ::7) Check pH again and adjust if necessary with small amounts of 5 N KOH | ||
::8) | ::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) | ::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) | ::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 [[#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. | ||
[[Image:B-Bioblast.png|left|30px|link=http://www.bioblast.at/index.php/Bioblast:About|Bioblast wiki]] | |||
== Popular Bioblast page == | == Popular Bioblast page == | ||
[[MiPNet14.13 Medium-MiR06]] has been accessed more than | [[MiPNet14.13 Medium-MiR06]] has been accessed more than | ||
:* 5,000 times (2013-09-07) | :* 5,000 times (2013-09-07) | ||
Revision as of 11:41, 27 April 2014
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.
Popular Bioblast page
MiPNet14.13 Medium-MiR06 has been accessed more than
- 5,000 times (2013-09-07)
