Daniel:Protocols/CProbeLigate
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C Probe Hybridization
This page describes the protocol for hybridizing C probes as used in PLAYR Design (as from Nolan lab).
Protocol
Resuspend oligos in nuclease free water to concentration of 100 uM
- Fix cells
- Pellet cells at 600g for 3 min and resuspend at concentration of ~1,000,000/mL
- Fix cells in medium with 1.6% paraformaldehyde for 10 min at RT
- Wash cells with wash buffer
- Pellet cells and permeabilize with ice-cold methanol for 10 min on ice Once in methanol cells may be stored long term
- Hybridization
Wash Buffer
- PBS
- 0.1% Tween
- 4 U/mL RNasin