Difference: ProceduresPolyacrylamideGelElectrophoresis (6 vs. 7)

Revision 72012-06-01 - JeffreyBarrick

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META TOPICPARENT name="ProtocolList"
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Polyacrylamide Gel Electrophoresis

Our gel rigs and supplies are from CBS Scientific.

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Spacer Width Gel Height Gel Width Gel Vol TEMED Vol 10% APS Vol
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1 mm 28 cm 16.5 cm 50 ml 50 Ál 400 Ál
 
1 mm 14.5 cm 16.5 cm 25 ml 25 Ál 200 Ál
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1 mm 28 cm 16.5 cm 50 ml 50 Ál 400 Ál
1.5 mm 14.5 cm 16.5 cm 30 ml 30 Ál 240 Ál
1.5 mm 28 cm 16.5 cm 75 ml 75 Ál 600 Ál
  Add 10 Ál of TEMED per 10 ml volume, then 50 Ál of 10% w/v APS per 10 ml volume to initiate polymerization of the gel. Pour it quickly into the glass plates with spacers clamped between them.
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Running the Gel

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You will need a high voltage power supply to run the large vertical polyacrylamide gels. Generally denaturing gels are run at a constant electrical power (Watts). This maintains a certain heated gel temperature during the run. For the 16.5 cm x 28.5 cm gels, use 25 W. Using a higher voltage can cause excessive heating that will crack the glass plates.
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You will need a high voltage power supply to run the large vertical polyacrylamide gels. Generally denaturing gels are run at a constant electrical power (Watts). This maintains a certain heated gel temperature during the run. For the 16.5 cm x 28.5 cm gels, use 35 W. Using a higher voltage can cause excessive heating that will crack the glass plates.
 

Reagents

 
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