Methods: Difference between revisions
(New page: {{Template:Rainey}} '''Staining a SDS-PAGE gel with coomassie blue'''<br> <br> '''1)''' Cover the gel with coomassie staining solution and leave on a shaker at slow speed for 30 minutes<br...) |
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'''5)''' When finished destaining, leave the gel in water. If desired preserve the gel in cellophane. | '''5)''' When finished destaining, leave the gel in water. If desired preserve the gel in cellophane. | ||
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'''1)''' After electrophoresis place the gel into the staining solution for 15 minutes with mild shaking<br> | '''1)''' After electrophoresis place the gel into the staining solution for 15 minutes with mild shaking<br> | ||
'''2)''' Place the gel into water and visualize under a UV lamp<br> | '''2)''' Place the gel into water and visualize under a UV lamp<br> | ||
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'''Running whole ''E. coli'' cells on an SDS-PAGE gel'''<br> | |||
'''1)''' Pellet 500 μL of culture in a micro centrifuge tube at maximum speed for 2 minutes<br> | |||
'''2)''' Remove the supernatant and add 50-250μL of 1x loading buffer with 50mM Dithiothreitol to the samples, depending on the size of the pellet. <br> | |||
'''3)''' Resuspend the pellet in the loading buffer, boil for 10 minutes<br> | |||
'''4)''' Place the microcentrifuge tubes on ice for a minute, then centrifuge at maximum speed for 2 minutes<br> | |||
'''5)''' Load 5μL from the '''top''' of the sample to your SDS-PAGE gel and enjoy. | |||
Revision as of 03:56, 13 March 2009
Staining a SDS-PAGE gel with coomassie blue
1) Cover the gel with coomassie staining solution and leave on a shaker at slow speed for 30 minutes
2) Remove the coomassie stain (can be reused), and replace with destaining solution for 30 minutes
3) Remove the destaining solution (into waste bottle) and replace with destaining solution for 30 minutes-overnight
4) If desired, repeat the destaining process
5) When finished destaining, leave the gel in water. If desired preserve the gel in cellophane.
Staining an agarose gel with ethidium bromide
1) After electrophoresis place the gel into the staining solution for 15 minutes with mild shaking
2) Place the gel into water and visualize under a UV lamp
Running whole E. coli cells on an SDS-PAGE gel
1) Pellet 500 μL of culture in a micro centrifuge tube at maximum speed for 2 minutes
2) Remove the supernatant and add 50-250μL of 1x loading buffer with 50mM Dithiothreitol to the samples, depending on the size of the pellet.
3) Resuspend the pellet in the loading buffer, boil for 10 minutes
4) Place the microcentrifuge tubes on ice for a minute, then centrifuge at maximum speed for 2 minutes
5) Load 5μL from the top of the sample to your SDS-PAGE gel and enjoy.
