Pouring Acrylamide Gels for Manual Run
 
Hazardous materials:
Acrylamide: neurotoxin




TEMED: corrosive
 For complete information, refer to the MSDS for each substance.
 
PPE required:
Gloves: Nitrile
Eyewear
Lab Coat
Closed toed shoes
 
Precautions to observe:
Mix polyacrylamide under hood
Avoid contact with skin/eyes
 
In case of contact:
If in contact with skin: remove clothing and use emergency shower or eye wash for 15 min
Call EHS to clean up water in hallway
 
Procedure:
 
Prepare plates (use the gel-pouring bench on the west side of the main lab)
1. Cut bench paper large enough to cover the surface that you will be using.
2. Choose 1 short and 1 long plate per gel
3. Set plates on white, plastic tube racks.
4. Clean the inside of both plates with 95% ethanol and a Kimwipe
5. Clean with isopropanol and a kimwipe.
 The plates should be completely free of smears or grease. After the isopropanol wash the glass 
 will feel kind of slippery.
6. Wipe a small amount of Rain-X on the inside of the short plate ONLY.
7. Place spacers on each (long) side of the long plate.
8. Place the clean, Rain-Xed side of the short plate on top of the spacers, facing the long plate.
 The bottom of the plates should be flush and spacers should be sandwiched inside the plates, 
 aligned with the outer, long edges.
9. Use 2-4 binding clips to clip and hold the plates together. 
10. Apply a casting boot to the bottom and sides of the plates removing clips as necessary.
 Make sure the short plate is on top of the long plate and the hole to insert the syringe at the 
 bottom of the boot is facing up.
 
Preparation of polyacrylamide (must be done in the hood)
1. For each gel to be poured use the following:
- 30ml 5% acrylamide (from 4 degree refrigerator)
- 90μl TEMED (in the hood)
- 90μl 10% APS (in 4 degree refrigerator)
2. Swirl gently in beaker.
 
Pouring polyacrylamide into gel
1. Carefully draw ~35ml of the polyacrylamide solution into a plastic syringe avoiding air bubbles.
2. Insert syringe into the hole in the bottom of the casting boot.
3. Slowly depress plunger, allowing acrylamide to flow in between the pates.
4. Proceed until the acrylamide fills the space between the plates and begins to run out at the top of the short plate.
5. Remove syringe.
6. Plug hole in casting boot with modified pipette tip.
7. Insert comb, blunt side down, into the space at between the plates at the top of the gel.
8. Use 1-3 binding clips to secure the comb and the top of the plates together.
9. Wait ~20 minutes to allow adequate polymerization.
 
Notes:
- If you have trouble and have to discard the gel, POLYMERIZE the acrylamide first, before disposal. NEVER pour unpolymerized acrylamide down the drain.
- Dispose of used bench paper after each use.
- If supplies are running low, be sure to notify the ordering or stocks personnel.
- Polymerized gels can be stored for future use by placing dampened (with water) paper towels over the top/bottom ends and covering the whole gel in saran wrap to avoid dehydration. Remove the casing boot prior to wrapping and storage.