Objective
Make a protein gel.
Description
1st Attempt at Making Gel Today
- Make the resolving gel:
- 1.65mL H2O
- 2.0mL 30% acrylamide mix
- 1.25mL 1.5M Tris (pH 8.8)
- 0.05mL 10% SDS
- 0.05mL 10% ammonium persulfate (add this right before pouring the gel)
- 0.002mL TEMED (add this right before pouring the gel)
- Pour the gel between the short plate and spacer plate (after they have been secured in the casting frame on the casting stand). Squirt a layer of methanol on top of the resolving gel for a straight, level layer of gel.
- This layer of gel never polymerized, so I need to try again.
2nd Attempt at Making Gel Today
- Make fresh 10% APS (0.1g ammonium persulfate + 1mL distilled H2O)
- Make new 200mL 1.5M Tris, pH 8.8 (36.34g Tris base + H2O + HCl (to pH))
- Make the resolving gel:
- 1.65mL H2O
- 2.0mL 30% acrylamide mix
- 1.25mL 1.5M Tris (pH 8.8)
- 0.05mL 10% SDS
- 0.05mL 10% ammonium persulfate (add this right before pouring the gel)
- 0.002mL TEMED (add this right before pouring the gel)
- Pour the gel between the short plate and spacer plate (after they have been secured in the casting frame on the casting stand). Squirt a layer of methanol on top of the resolving gel for a straight, level layer of gel.
Notes
Neither resolving gel solidified today. I will have to continue to troubleshoot this problem tomorrow.
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