User:Linh N Le/Notebook/2009/08/26: Difference between revisions
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***Ideally the LN2 level should be such that the liquid is splashing the top of the racks | ***Ideally the LN2 level should be such that the liquid is splashing the top of the racks | ||
*Also have an Ice bucket with ice ready so that when you take out the tubulin to mix and aliquot out you can keep it on ice | *Also have an Ice bucket with ice ready so that when you take out the tubulin to mix and aliquot out you can keep it on ice | ||
*The manuals state that both of the tubulins must have a concentration of 10mg/ml to work | |||
===Rhodamine Tubulin=== | ===Rhodamine Tubulin=== | ||
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===Unlabeled Tubulin=== | ===Unlabeled Tubulin=== | ||
*Same as above but we made 38, 5ul aliquots in the 500ul tubes | *Same as above but we made 38, 5ul aliquots in the 500ul tubes | ||
*We added 200ul GPEM+cushion (180ul GPEM + 20ul cushion) to a 1mg tube of the tubulin (notice the higher concentration of the unlabeled kind, hence more aliquots were made), mixed it and put 5ul into a 500ul tube, then flash froze | *We added 200ul of ''cold'' GPEM+cushion (180ul GPEM + 20ul cushion) to a 1mg tube of the tubulin (notice the higher concentration of the unlabeled kind, hence more aliquots were made), mixed it and put 5ul into a 500ul tube, then flash froze | ||
*For this, we used the red icebucket and 5 racks to fit all the tubes into the LN2 | *For this, we used the red icebucket and 5 racks to fit all the tubes into the LN2 | ||
*The tubes were then placed into a 50ml falcon tube for storage in the -80C | |||
Revision as of 13:51, 26 August 2009
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To Do
Tubulin
ProcedureNote: It is important to keep the tubulin cold, so prepare all the tubes, buffers and LN2 before you start
Rhodamine Tubulin
Unlabeled Tubulin
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