Daniel:Notebook/RNAFISH/2015-1-29: Difference between revisions
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>Djacobse (Created page with "=Oligo Set Preparations= Back to Calendar These are the oligos that I ordered on 08/04/2014. Now that I am...") |
>Djacobse |
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#Add 384 uL nfH2O, diluting 5:1, for final concentration of 200 uM | #Add 384 uL nfH2O, diluting 5:1, for final concentration of 200 uM | ||
#Check concentration in nanodrop at 1:1, 5:1, and 10:1 dilutions for accuracy | #Check concentration in nanodrop at 1:1, 5:1, and 10:1 dilutions for accuracy | ||
==Nanodrop Results== | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt" | |||
| align="center" width="65" height="40" valign="bottom" | | |||
|style="font-weight:bold" width="65" align="center" | 1:1 ng/uL | |||
|style="font-weight:bold" width="65" align="center" | 5:1 ng/uL | |||
|style="font-weight:bold" width="65" align="center" | 10:1 ng/uL | |||
|style="font-weight:bold" width="72" align="center" | Average (ng/uL) | |||
|style="font-weight:bold" width="65" align="center" | STDev (ng/uL) | |||
|- style="font-size:12pt" valign="bottom" | |||
|style="font-weight:bold" height="15" | FOXP2 | |||
| align="center" | 959.3 | |||
| align="center" | 194.2 | |||
| align="center" | 97.5 | |||
| align="center" | 968.4 | |||
| align="center" | 8.2 | |||
|- style="font-size:12pt" valign="bottom" | |||
|style="font-weight:bold" height="15" | GAD1 | |||
| align="center" | 951.9 | |||
| align="center" | 190 | |||
| align="center" | 94.7 | |||
| align="center" | 949.6 | |||
| align="center" | 2.5 | |||
|} | |||
So they are both just under 1 ug/uL. |
Latest revision as of 23:47, 29 January 2015
Oligo Set Preparations[edit]
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)
- Store the remaining oligos at -20C
- Add 384 uL nfH2O, diluting 5:1, for final concentration of 200 uM
- Check concentration in nanodrop at 1:1, 5:1, and 10:1 dilutions for accuracy
Nanodrop Results[edit]
1:1 ng/uL | 5:1 ng/uL | 10:1 ng/uL | Average (ng/uL) | STDev (ng/uL) | |
FOXP2 | 959.3 | 194.2 | 97.5 | 968.4 | 8.2 |
GAD1 | 951.9 | 190 | 94.7 | 949.6 | 2.5 |
So they are both just under 1 ug/uL.