Daniel:Notebook/RNAFACS/2016-1-11: Difference between revisions

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>Djacobse
(Created page with "=Testing Poly-T Primers= Back to Calendar ==Buffers== *Wash Buffer A **200 uL Stellaris RNA FISH Wash Buffer A (Biosearch Technologies Cat# SMF-...")
 
>Djacobse
 
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**Mix well by vortexing gently
**Mix well by vortexing gently
*Hybridization Buffer
*Hybridization Buffer
**900 μL Stellaris RNA FISH Hybridization Buffer
**90 μL Stellaris RNA FISH Hybridization Buffer
**100 μL Deionized Formamide
**10 μL Deionized Formamide


===Protocol===
===Protocol===

Latest revision as of 22:51, 11 January 2016

Testing Poly-T Primers[edit]

Back to Calendar

Buffers[edit]

  • Wash Buffer A
    • 200 uL Stellaris RNA FISH Wash Buffer A (Biosearch Technologies Cat# SMF-WA1-60)
    • Add 700 uL Nuclease-free water
    • Add 100 uL Deionized Formamide
    • Mix well by vortexing gently
  • Hybridization Buffer
    • 90 μL Stellaris RNA FISH Hybridization Buffer
    • 10 μL Deionized Formamide

Protocol[edit]

  1. Aspirate the 70% ethanol
  2. Add 1 mL of Wash Buffer A (see recipe above), and incubate at room temperature for 2-5 minutes
  3. Add 1 uL of probe stock solution to 100 uL Hybridization Buffer for working concentration of 125 nM
  4. Aspirate off Wash Buffer A
  5. Dispense 100 μL of the Hybridization Buffer onto the center of the dish
  6. Incubate in the dark at 37 °C for at least 4 hours. (Incubation can be continued up to 16 hours)