Daniel:Notebook/RNAFACS/2015-11-20

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Revision as of 15:54, 20 November 2015 by >Djacobse (→‎Day 1-Silane Binding)
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PAGE Gel Creation[edit]

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The reagents finally came in for making gels on slides. Today I'll start with protocol #1.

Day 1-Silane Binding[edit]

  1. Load slides (facing the same direction) into metal slide racks
  2. Sterilize the following items (15 minutes under UV light in PCR hood):
    1. two large glass Coplin buckets & lids
    2. metal slide racks with slides loaded
    3. 1 L dH20 in a plastic container
    4. stir bar
  3. Add 220 uL Acetic Acid to the 1 L dH20 (pH -> 3.5; no need to measure)
  4. Add 4 mL Bind-Silane
  5. Mix for 15 minutes using stir-bar.
  6. Place loaded slide racks in glass Coplin buckets
  7. Add stirred mixture to Coplin buckets (500 mL per bucket; enough to cover the slides)
  8. Cover buckets and incubate with slow shaking for 1 hour.
  9. Leaving racks in buckets, rinse 3 times with dH20
  10. Rinse once with 100% ethanol.
  11. Remove slide-racks from buckets and allow slides to dry (in racks) under AirClean hood.
  12. Transfer slides to plastic storage boxes (keep the Teflon coated sides of the slides facing in the same direction)
  13. Label boxes with initals and date of treatment.
  14. Place slide boxes in dessicator for long-term storage.