Daniel:Notebook/RNAFACS/2015-11-20
Jump to navigation
Jump to search
PAGE Gel Creation[edit]
The reagents finally came in for making gels on slides. Today I'll start with protocol #1.
Day 1-Silane Binding[edit]
- Load slides (facing the same direction) into metal slide racks
- Sterilize the following items (15 minutes under UV light in PCR hood):
- two large glass Coplin buckets & lids
- metal slide racks with slides loaded
- 1 L dH20 in a plastic container
- stir bar
- Add 220 uL Acetic Acid to the 1 L dH20 (pH -> 3.5; no need to measure)
- Add 4 mL Bind-Silane
- Mix for 15 minutes using stir-bar.
- Place loaded slide racks in glass Coplin buckets
- Add stirred mixture to Coplin buckets (500 mL per bucket; enough to cover the slides)
- Cover buckets and incubate with slow shaking for 1 hour.
- Leaving racks in buckets, rinse 3 times with dH20
- Rinse once with 100% ethanol.
- Remove slide-racks from buckets and allow slides to dry (in racks) under AirClean hood.
- Transfer slides to plastic storage boxes (keep the Teflon coated sides of the slides facing in the same direction)
- Label boxes with initals and date of treatment.
- Place slide boxes in dessicator for long-term storage.