Hosuk:LabNotes/2013-10-28: Difference between revisions
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>Hosuki78 (Created page with "*LabNote ===Detect RAB7A, and compare to ACTB=== *Use 10/22 sample (MatTek dish, S2) *RAB7A : Resuspension --> 200uM *1uL 200uM RAB7A + 2uL Cy5 1st rolon...") |
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*1uL 200uM ACTB + 2uL Cy5 1st rolony adapter in 197 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 | *1uL 200uM RAB7A + 2uL Cy5 1st rolony adapter in 197 uL 2x SSC --> 2 tubes | ||
*2mL 80% Formamide | |||
*2x SSC, PBS | |||
Revision as of 17:31, 28 October 2013
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
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+1st only), image --> 1st round ACTB only
- Strip, Add 100uL (RAB7A+1st only), image --> 2nd round RAB7A only
- Strip, Add 100uL (ACTB+1st only), image --> 2nd round ACTB only