IGEM:Peking/2007/Count:Agarose Gel Electrophoresis: Difference between revisions

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(New page: ==Casting agarose gels== ===Materials=== *gel box, gel tray, comb *Agarose *''GeneFinder'' safe dye *1X TAE *Microwave *100 ml glass bottle or beaker ===Procedure=== *Assemble casting...)
 
 
(5 intermediate revisions by the same user not shown)
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==Casting agarose gels==
==Casting agarose gels==
===Materials===
===Materials===
*gel box, gel tray, comb  
*Gel box, Gel tray, Gel comb  
*Agarose  
*Agarose  
*''GeneFinder'' safe dye
*''GeneFinder'' safe dye(200X stock)
*1X TAE  
*1X TAE  
*Microwave  
*Microwave  
Line 9: Line 9:


===Procedure===
===Procedure===
*Assemble casting trays ,gel box and gel comb,choose the comb of appropriate width, size, and number into niche at end of the tray.  
#Assemble casting trays ,gel box and gel comb,choose the comb of appropriate width, size, and number into niche at end of the tray.  
*agarose solution(1% gel)：0.2g agarose + 20ml 1X TAE for small gels or 0.4g agarose + 40ml 1X TAE for large gels.
#agarose solution(1% gel)：0.2g agarose + 20ml 1X TAE for small gels or 0.4g agarose + 40ml 1X TAE for large gels.
#add agarose solution in a 100ml bottle or beaker.
#Microwave bottle for 1min
#Remove from microwave and let cool
#Once gel is cooled so that it can be touched comfortably with your gloved hand, add 50 μL(for small gel) or 100 μL(for large gels) ''GeneFinder'' safe dye.
#Pour gel into casting trays.
==prepare Loading dye==
*mix 10X loading buffer and 200X ''GeneFinder'' safe dye in a 1:1 ratio.
*Store at 4°C
==Running agarose gels==
#Place the gel in gel box.
#Add enough 1X TAE to fill the reservoirs at both ends of the gel box and cover the surface of the gel--the gel should be immersed.
#Mix loading dye with samples:Typically, use 0.5 μL loading dye per 2 μL of sample.
#Load samples left to right.
#Place gel box cover on gel box such that your samples will run towards the positive, red electrode. Make sure that the cables from the cover are connected to the power supply correctly.
#Turn on the power supply and run your gel at ~100 V for 13~15 mins.
#Verify that bubbles are rising from the electrodes once you start your gel to ensure your gel is running properly.  


Pour gel into casting trays.
==Marker standard==
*We use ''transgen'' marker.
*standard marker 1
[[Image:Peking_2007_Marker_I.JPG‎]]
*DL2000 plus marker
[[Image:Peking_2007_DL2000_plus.JPG‎]]
 
==See also==
* [[Agarose_gel_electrophoresis]]
* [[Endy:Agarose gel]]
* [[Knight:Agarose gel electrophoresis]]

Latest revision as of 07:02, 6 August 2007

Casting agarose gels

Materials

  • Gel box, Gel tray, Gel comb
  • Agarose
  • GeneFinder safe dye(200X stock)
  • 1X TAE
  • Microwave
  • 100 ml glass bottle or beaker

Procedure

  1. Assemble casting trays ,gel box and gel comb,choose the comb of appropriate width, size, and number into niche at end of the tray.
  2. agarose solution(1% gel)：0.2g agarose + 20ml 1X TAE for small gels or 0.4g agarose + 40ml 1X TAE for large gels.
  3. add agarose solution in a 100ml bottle or beaker.
  4. Microwave bottle for 1min
  5. Remove from microwave and let cool
  6. Once gel is cooled so that it can be touched comfortably with your gloved hand, add 50 μL(for small gel) or 100 μL(for large gels) GeneFinder safe dye.
  7. Pour gel into casting trays.

prepare Loading dye

  • mix 10X loading buffer and 200X GeneFinder safe dye in a 1:1 ratio.
  • Store at 4°C

Running agarose gels

  1. Place the gel in gel box.
  2. Add enough 1X TAE to fill the reservoirs at both ends of the gel box and cover the surface of the gel--the gel should be immersed.
  3. Mix loading dye with samples:Typically, use 0.5 μL loading dye per 2 μL of sample.
  4. Load samples left to right.
  5. Place gel box cover on gel box such that your samples will run towards the positive, red electrode. Make sure that the cables from the cover are connected to the power supply correctly.
  6. Turn on the power supply and run your gel at ~100 V for 13~15 mins.
  7. Verify that bubbles are rising from the electrodes once you start your gel to ensure your gel is running properly.

Marker standard

  • We use transgen marker.
  • standard marker 1

  • DL2000 plus marker

See also

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