Hosuk:LabNotes/2013-10-28
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Detect RAB7A, and compare to ACTB
- Use 10/22 sample (MatTek dish, S2)
- RAB7A : Resuspension --> 200uM
- 1uL 200uM RAB7A + 2uL Cy5 1st rolony adapter in 197 uL 2x SSC
Preparation
- 1uL 200uM ACTB + 199 uL 2x SSC --> 2 tubes
- 1uL 200uM RAB7A + 199 uL 2x SSC --> 2 tubes
- 1uL 200uM ACTB + 2uL Cy5 1st rolony adapter in 197 uL 2x SSC --> 2 tubes
- 1uL 200uM RAB7A + 2uL Cy5 1st rolony adapter in 197 uL 2x SSC --> 2 tubes
- 2mL 80% Formamide
- 2x SSC, PBS
- Aspiration setup, vacuum pump
Procedure
- Strip, Add 100uL (RAB7A+1st rolony probe mix), image --> 1st round RAB7A
- Strip, Add 100uL (ACTB+1st rolony probe mix), image --> 1st round ACTB
- Strip, Add 100uL (RAB7A+1st rolony probe mix), image --> 2nd round RAB7A
- Strip, Add 100uL (ACTB+1st rolony probe mix), image --> 2nd round ACTB
- Strip, Add 100uL (RAB7A only), image --> 1st round RAB7A only
- Strip, Add 100uL (ACTB only), image --> 1st round ACTB only
- Strip, Add 100uL (RAB7A only), image --> 2nd round RAB7A only
- Strip, Add 100uL (ACTB only), image --> 2nd round ACTB only