Team:TU Darmstadt/protocols/SDS-Page

(Difference between revisions)

Revision as of 18:53, 4 October 2013

Equipment

Chemicals & consumables

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

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

@@ Line 218: / Line 218: @@
 <p text-aligne:left style="margin-left:60px; margin-right:50px"><font size="3" color="#F0F8FF" face="Arial regular">
 <br>
-<B>Buffers<br></B>
+<B>Buffers & gels<br></B>
 <div align="left" style="margin-left:60px; margin-right:50px">
 <ul>
-<li class=list1>- Separation buffer (18.17 g Tris, 0.40 g SDS, 90 ml Aqua dest., pH = 8.8)</li>
+<li class=list1>- Separation buffer (0.5 M Tris, 0.4% g SDS, pH = 8.8)</li>
-<li class=list1>- Rotiphorese® (30%)</li>
+<li class=list1>- Stacking buffer (0.5 M Tris, 0.4% g SDS, pH = 6.6)</li>
-<li class=list1>- Tris HCl</li>
+<li class=list1>- Running buffer (0.25 M Tris, 2 M Glycin, 1% SDS , pH = 8.3)</li>
-<li class=list1>- Glycine</li>
+<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>- APS</li>