Difference between revisions of "Team:Aachen/Notebook/Documentation/Glycogen Characterization"
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* experiment should work if the feed is treated with HCl for acid hydrolysis first | * experiment should work if the feed is treated with HCl for acid hydrolysis first | ||
− | * Dinitrosalicylic acid staining | + | * Dinitrosalicylic acid staining (OD of WT, ΔglgX and ΔglgP were adjusted) |
− | * | + | * 1:100, 1:50, 1:25, 1:12.5, 1:6.25, 1:3.125, and 1:1.5625 dilutions of WT, ΔglgP, and ΔglgX were prepared; 50 µL sample volume. The same volume of Dinitrosalicylic acid was added; total volume was 100 µL. |
+ | * blank: 1:1 dilution of dinitrosalicylic acid in water. | ||
+ | * the stained samples were heated for 10 minutes at 90 °C | ||
+ | * the heated samples were cooled down in a 25 °C heater | ||
+ | * samples were analyzed in plate reader at 540 nm | ||
==15-09-13== | ==15-09-13== | ||
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* two 10 ml LB overnight cultures of #E4QF# were inoculated | * two 10 ml LB overnight cultures of #E4QF# were inoculated | ||
* no difference in growth or glycogen production could be observed | * no difference in growth or glycogen production could be observed | ||
+ | |||
+ | * Dinitrosalicylic acid staining | ||
+ | ** two replicates of 5 mL LB overnight cultures of WT and ΔglgB were prepared | ||
==15-09-14== | ==15-09-14== | ||
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**MeOh was added at t: 5 (OD 0.769 - 0.802) at 3:15 p.m. | **MeOh was added at t: 5 (OD 0.769 - 0.802) at 3:15 p.m. | ||
* no influence of MeOH could be observed | * no influence of MeOH could be observed | ||
+ | |||
+ | * Dinitrosalicylic acid staining | ||
+ | ** 3 biological replicates of ''glg B'' strain and wild type were prepared by transferring 2 mL of LB overnight cultures to M9 Nitrogen limitation media. The calculated starting OD was 0.1. | ||
+ | |||
+ | ==15-09-15== | ||
+ | * Dinitrosalicylic acid staining | ||
+ | ** M9 N-limitation cultures were [[Team:Aachen:purified | ||
==15-09-16== | ==15-09-16== |
Revision as of 03:32, 19 September 2015