Matt:LabNotes/2015-6-15: Difference between revisions
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| align="center" style="background:#f0f0f0;"|'''Stdev''' | | align="center" style="background:#f0f0f0;"|'''Stdev''' | ||
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| KIT 1:1|| | | KIT 1:1||1053.5||1053.5|||| | ||
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| KIT 1:5|| | | KIT 1:5||212.5||1062.5|||| | ||
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| KIT 1:10|| | | KIT 1:10||104.9||1049||1055||6.87 | ||
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| SNAP25 1:1|| | | SNAP25 1:1||1039.5||1039.5|||| | ||
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| SNAP25 1:5|| | | SNAP25 1:5||207.7||1038.5|||| | ||
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| SNAP25 1:10|| | | SNAP25 1:10||107.4||1074||1050.67||20.21 | ||
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Revision as of 01:36, 16 June 2015
Motor Neurons RNA FISH Dye Coupling
- Previous RNA FISH attempt detected no signal
- Try again with higher expressed genes (CUX2 and KIT)
Probe Resuspension
- Table of ordered plates
- KIT: Plate=KIT-SNAP25-20150128 Wells=A1-D12
- SNAP25: Plate=KIT-SNAP25-20150128 Wells=E1-H12
- Table of ordered plates
- Resuspend each oligo in 15 uL nuclase-free H2O (1 mM)
- Take 2 uL from each oligo well and combine them in a single 1.5 mL tube (total concentration 1 mM)
- Store the remaining oligos at -20C
- Add 450 uL nuclase-free, diluting 10:1, for final concentration of 100 uM
- Technically closer to 5:1 dilution but final result is closer to 10:1 because starting concentration is actually lower...
- Goal is to get 1ug/ul DNA concentration
- Check concentration in nanodrop at 1:1, 5:1, and 10:1 dilutions for accuracy