Pouring Acrylamide Gels for the Basestation
 
Hazardous materials:
Acrylamide: neurotoxin

TEMED: corrosive
Photoinitiator: ??
 For complete information, refer to the MSDS for each substance.
 
PPE required:
Gloves: Nitrile
Eyewear
Lab Coat
Closed toed shoes
 
Precautions to observe:
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
 
 
Preparing Plates:
1. Choose a top and bottom plate corresponding to the correct size (20cm or 30cm) and number of samples you wish to run (50l or 100).
2. Place the outside of the plates on white plastic tub racks, so the inside of the plates is facing upward.
For the bottom plate (without the buffer chambers), look for arrows on the side of the plate. The arrows should always face upward and indicate the inside of the plate.
3. Clean plates with nanopure water and a Kimwipe.
4. Then clean plates again with Ethanol and Kimwipes. Be sure the plates are completely dry.
5. Place the bottom plate (without the buffer chambers) in the MJ casting stand. Push plate flush toward the top, right corner of the stand.
6. Choose a mylar spacer of appropriate size for your plates (20cm or 30cm). The spacer should not be torn or wrinkled.
7. Center the spacer on top of the plate, aligning it with the corners.
8. Gently place the upper plate on top of the bottom plate and spacers. The wells should be away from the clamps on the casting stand. Both plates should be pushed tight and flush against the top, right corner of the casting stand.
9. Use four binding clips to hold the plates together.
 
Gel Pouring:
1. Place the o-ring in the loading end of the top plate. Be sure the o-ring is dry and properly seated.
2. Using the clamps on the casting stand, clip the o-ring in place.
3. Choose the proper comb for your gel. Make sure the screws are recessed into the bracket so the comb can be fully inserted.
4. With a 25ml syringe, carefully draw up ~15ml of acrylamide, avoiding bubbles.
5. Insert syringe in anode seal and slowly depress plunger, injecting acryalmide between the plates.
6. Continue injection until acrylamide the wells on the opposite end and flows up out of all of the wells.
7. Insert the comb, being careful to keep the comb level during insertion to avoid breaking it. Push comb down so sides sit flush against the bottom plate.
8. Unclamp anode seal and place o-ring and syringe in a waste bucket until it can be cleaned.
 
Gel Polymerization
1. Pre-run the UV box before polymerization. During pre-run, check to insure all light bulbs are working.
2. Remove the gel assembly from the casting stand (keeping binding clips in place) and transfer to the UV box.
3. Clean any acrylamide from the casting stand using Kimwipes and nanopure water. Place any Kimwipes used on the uvbox.
4. Close the lid and press start on the timer.
5. After the first polymerization cycle, the Kimwipes may be discarded.
6. Turn the gel 180 degrees, close the lid, and run another 5 minute polymerization cycle. This may be repeated again if desired.
7. Place gel back on casting stand and remove binder clips.
8. Leaving the comb in place, squirt around the comb, and in the cathode chamber to remove unwanted aryclamide.
9. Use the the vacuum to remove all acrylamide and water from the chamber.
10. Clean the top of the plate with nanopure water.