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" | &nbsp;
|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]

Back to Calendar

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.

  1. Resuspend each oligo in 15 uL nfH2O (1 mM)
  2. Take 2 uL from each oligo well and combine them in a single 1.5 mL tube (total concentration 1 mM)
    1. Store the remaining oligos at -20C
  3. Add 384 uL nfH2O, diluting 5:1, for final concentration of 200 uM
  4. 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.