Daniel:Notebook/RNAFISH/2014-8-30
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Trial Run (Started 08/29/2014)
Tissue Section Preparation (From 08/29/2014)
- Wash the tissue in PBS, 3 × 30 min
- Incubate the tissue in a sucrose series with increasing concentrations:
- 10% for 1 h
- 20% for 1 h
- 30% at 4C overnight
Oligo Set Preparations
These are the oligos that I ordered on 08/04/2014. Now that I am ready to do the experiment, I need to prep them. First thing is to resuspend the oligos. By order, each oligo has 15 nmol.
- Resuspend each oligo in 15 uL nfH2O (1 mM)
- Take 2 uL from each oligo well and combine them in a single 1.5 mL tube (total concentration 1 mM)
- Add 432 uL nfH2O, diluting 10:1, for final concentration of 100 uM
- Check concentration in nanodrop at 1:1, 5:1, and 10:1 dilutions for accuracy
Nanodrop Results
Sample | 1:1 Measured | 5:1 Measured | 5:1 Corrected | 10:1 Measured | 10:1 Corrected | Average | Standard Deviation |
GAD1 | 558 | 109.3 | 546.5 | 43.2 | 432 | 512.2 | 69.7 |
FOXP2 | 554.3 | 114.9 | 574.5 | 54.5 | 545 | 557.9 | 15.1 |
RBFOX3 | 558.9 | 115.2 | 576 | 56.8 | 568 | 567.6 | 8.6 |
CUX2 | 582.2 | 124.9 | 624.5 | 60.6 | 606 | 604.2 | 21.2 |
So each one has around 0.5 ug/uL. Pretty good.