Daniel:Notebook/HiResChrPaint/2014-7-18: Difference between revisions
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##Total volume will be 64 uL | ##Total volume will be 64 uL | ||
#Add 742.4 uL nfH2O, dilutes to 1 ug/uL total pool | #Add 742.4 uL nfH2O, dilutes to 1 ug/uL total pool | ||
##Create dilution series and measure in nanodrop | |||
===Nanodrop Results=== | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt;font-weight:bold" align="center" valign="bottom" | |||
| width="78" height="15" | Dilution (X:1) | |||
| width="65" | ng/uL | |||
| width="65" | proj. ug/uL | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| align="center" height="15" | 5 | |||
| align="center" | 79.6 | |||
| align="center" | 0.40 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| align="center" height="15" | 10 | |||
| align="center" | 38.2 | |||
| align="center" | 0.38 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| align="center" height="15" | 20 | |||
| align="center" | 19.3 | |||
| align="center" | 0.39 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| align="center" height="15" | 50 | |||
| align="center" | 7.4 | |||
| align="center" | 0.37 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
| align="center" height="15" | 100 | |||
| align="center" | 3.7 | |||
| align="center" | 0.37 | |||
|- style="font-size:12pt" align="center" valign="bottom" | |||
|style="font-weight:bold" height="15" | Average | |||
| align="center" | | |||
|style="font-weight:bold" align="center" | 0.38 | |||
|} | |||
So I guess my calculations were a bit off, but 0.4 ug/uL is pretty good for FISH, especially considering I need 30 pmol per experiment (0.4 ug/uL ~60 pmol/uL). | |||
[[Category:HRCP]] [[Category:20140718]] |
Latest revision as of 21:19, 19 July 2014
Dystrophin Probes[edit]
The dystrophin probes I designed last week arrived from IDT. Attached are their specs:
The oligos arrived as dry oligos in a plate. The calculations I made are in the following text file:
Oligo Pool Synthesis[edit]
- Resuspend each oligo to make them all 1 mM (See Plate Specs for values)
- Combine 2 uL from each oligo
- Makes 12.5 ug/uL total pool
- Total volume will be 64 uL
- Add 742.4 uL nfH2O, dilutes to 1 ug/uL total pool
- Create dilution series and measure in nanodrop
Nanodrop Results[edit]
Dilution (X:1) | ng/uL | proj. ug/uL |
5 | 79.6 | 0.40 |
10 | 38.2 | 0.38 |
20 | 19.3 | 0.39 |
50 | 7.4 | 0.37 |
100 | 3.7 | 0.37 |
Average | 0.38 |
So I guess my calculations were a bit off, but 0.4 ug/uL is pretty good for FISH, especially considering I need 30 pmol per experiment (0.4 ug/uL ~60 pmol/uL).