All steps are performed in a darkroom using a red safe light.
1. Put a foil-wrapped glass sheet onto a tray of ice to cool in the darkroom.
2. Place about 15 mL of emulsion ribbons into a 50-mL Falcon tube, and incubate in a 42°C waterbath to melt them.
3. Decant 8 mL of the melted emulsion into a slide mailer, and replace at 42°C.
4. Prepare 8.8 mL water containing 180 pL glycerol, and add to the melted emulsion. Mix gently with a glass pipet to expel bubbles, and leave in water bath for 10 min to warm to 42°C.
5. Dip slides briefly and with a uniform speed into the emulsion, wipe the back (nonsection side) of each, and lay section side up on cold tray for 10 min to set. If bubbles or streaks are visible, then redip the section.
6. Place slides in a box for emulsion to harden in complete darkness for 2 h, then transfer to a sealed, light-tight box containing silica gel packets, and store at 4°C for a few days to 2 wk depending on abundance of target transcript.
Was this article helpful?