Daniel:Notebook/RNAFACS/2016-1-11
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Testing Poly-T Primers[edit]
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]
- Aspirate the 70% ethanol
- Add 1 mL of Wash Buffer A (see recipe above), and incubate at room temperature for 2-5 minutes
- Add 1 uL of probe stock solution to 100 uL Hybridization Buffer for working concentration of 125 nM
- Aspirate off Wash Buffer A
- Dispense 100 μL of the Hybridization Buffer onto the center of the dish
- Incubate in the dark at 37 °C for at least 4 hours. (Incubation can be continued up to 16 hours)