Team:Wageningen UR/Protocol/SDSPolyacrylamide

From 2012.igem.org

(Difference between revisions)
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*Wear gloves
*Wear gloves
*Clean glass slides with MQ and dry
*Clean glass slides with MQ and dry
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 +
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'''running gel'''
<ol>
<ol>
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<li>Places glass slides in holders and tighten in rack.</li>
<li>Places glass slides in holders and tighten in rack.</li>
polymerization:
polymerization:
-
<li>Add 4 ul TEMED to the running gel mix, mix and pipet the gel into the slipes until the green edge.</li>
+
<li>Add 4 ul TEMED to the running gel mix, mix and pipet the gel into the slides until the green edge.</li>
<li>Add a layer of iso-propanol on top of the polymerizing gel to equalize.</li>
<li>Add a layer of iso-propanol on top of the polymerizing gel to equalize.</li>
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</ol>
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 +
 +
'''Stacking gel'''
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 +
<ol>
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<li>When the running gel has fully polymerized, remove the remaing iso-propanol with a tissue</li>
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<li>Prepare stacking gel mix in a small beaker for 2 gels:</li>
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*1.4 ml MQ
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*0.33 ml 30% acrylamide mix
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*0.25 ml 1 M TRIS pH 6.8
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*0.02 ml 10% SDS
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*0.02 ml 10% ammonium persulphate
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<li>Add 2 ul TEMED to the stacking gel mix, mix and pipet the gel onto the running gel.</li>
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<li>Place comb</li>
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<li>After the stacking gel has fully polymerized, the gels can be stored at 4 degree Celcius in a box with wet tissues.</li>
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</ol>

Revision as of 11:35, 30 May 2012


SDS-polyacrylamide and native gel electrophoresis

Reagents & Materials

Procedure

  • Wear gloves
  • Clean glass slides with MQ and dry


running gel

  1. Prepare running gel mix in a small beaker for 2 gels:
    • 4 ml MQ
    • 3.3 ml 30% acrylamide mix
    • 2.5 ml 1.5 M TRIS pH 8.8
    • 0.1 ml 10% SDS
    • 0.1 ml 10% ammonium persulphate
  2. Places glass slides in holders and tighten in rack.

    3. polymerization:

  3. Add 4 ul TEMED to the running gel mix, mix and pipet the gel into the slides until the green edge.
  4. Add a layer of iso-propanol on top of the polymerizing gel to equalize.


Stacking gel

  1. When the running gel has fully polymerized, remove the remaing iso-propanol with a tissue
  2. Prepare stacking gel mix in a small beaker for 2 gels:
    • 1.4 ml MQ
    • 0.33 ml 30% acrylamide mix
    • 0.25 ml 1 M TRIS pH 6.8
    • 0.02 ml 10% SDS
    • 0.02 ml 10% ammonium persulphate
  3. Add 2 ul TEMED to the stacking gel mix, mix and pipet the gel onto the running gel.
  4. Place comb
  5. After the stacking gel has fully polymerized, the gels can be stored at 4 degree Celcius in a box with wet tissues.
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