Team:TU Darmstadt/protocols/SDS-Page

From 2013.igem.org

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<B>Buffers<br></B>
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<B>Buffers & gels<br></B>
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<li class=list1>- Separation buffer (18.17 g Tris, 0.40 g SDS, 90 ml Aqua dest., pH = 8.8)</li>
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<li class=list1>- Separation buffer (0.5 M Tris, 0.4% g SDS, pH = 8.8)</li>
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<li class=list1>- Rotiphorese® (30%)</li>
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<li class=list1>- Stacking buffer (0.5 M Tris, 0.4% g SDS, pH = 6.6)</li>
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<li class=list1>- Tris HCl</li>
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<li class=list1>- Running buffer (0.25 M Tris, 2 M Glycin, 1% SDS , pH = 8.3)</li>
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<li class=list1>- Glycine</li>
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<li class=list1>- Separation gel 12.5% (5 ml Separation buffer, 6.25 ml Rotiphorese®, 3.75 ml Aqua dest., 30 µl TEMED, 30 µl APS (40%))</li>
<li class=list1>- TEMED</li>
<li class=list1>- TEMED</li>
<li class=list1>- APS</li>
<li class=list1>- APS</li>

Revision as of 18:53, 4 October 2013







Labbook | Matierals | Protocols

Protocols


Equipment

  • - Two glass plates
  • - Gel caster
  • - Comb
  • - VWR Power Source 300V


Chemicals & consumables

  • - SDS
  • - Rotiphorese® (30%)
  • - Tris HCl
  • - Glycine
  • - TEMED
  • - APS
  • - Aqua dest.
  • - Isopropyle alcohol


Buffers & gels

  • - Separation buffer (0.5 M Tris, 0.4% g SDS, pH = 8.8)
  • - Stacking buffer (0.5 M Tris, 0.4% g SDS, pH = 6.6)
  • - Running buffer (0.25 M Tris, 2 M Glycin, 1% SDS , pH = 8.3)
  • - Separation gel 12.5% (5 ml Separation buffer, 6.25 ml Rotiphorese®, 3.75 ml Aqua dest., 30 µl TEMED, 30 µl APS (40%))
  • - TEMED
  • - APS
  • - Aqua dest.
  • - Isopropyle alcohol


Procedure

  1. prepare the separating gel and fill it into the chamber
  2. pour 1 mL isopropyle alcohol on the top of the gel to destroy air bubbles and prevent dehydration
  3. discard the isoprpyl alcohol and pour the prepared stacking gel
  4. stick in the comb
  5. store the gel in wet cloth (to prevent dehydration) at 4°C