Daniel:Notebook/HiResChrPaint/2014-9-19: Difference between revisions
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>Djacobse (Created page with "=FISH= Back to Calendar Today I'll perform the FISH experiments on the slides I prepared [[Daniel:Notebook/HiResChrPaint/2014-9-17|Wednesda...") |
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'''Sample Matrix''' | '''Sample Matrix''' | ||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="background-color:#CCC0DA;font-size:12pt;font-weight:bold" | |||
| width="65" height="33" valign="bottom" | Sample | |||
| width="65" align="center" valign="bottom" | [[Daniel:Notebook/RNAFISH/2014-8-31#Labeling Results|GAD1-488 (pmol)]] | |||
| width="65" align="center" valign="bottom" | [[Daniel:Notebook/RNAFISH/2014-8-31#Labeling Results|FOXP2-546 (pmol)]] | |||
| width="79" align="center" valign="bottom" | [[Daniel:Notebook/RNAFISH/2014-8-31#Labeling Results|RBFOX3-594 (pmol)]] | |||
| width="65" align="center" valign="bottom" | [[Daniel:Notebook/RNAFISH/2014-8-31#Labeling Results|CUX2-647 (pmol)]] | |||
| width="65" align="center" valign="bottom" | [[Daniel:Notebook/HiResChrPaint/2014-9-11|DMD-B-488 (pmol)]] | |||
| width="65" align="center" valign="bottom" | [[Daniel:Notebook/HiResChrPaint/2014-9-11|PPRL-A2-488 (pmol)]] | |||
| width="65" align="center" valign="bottom" | [[Daniel:Notebook/HiResChrPaint/2014-9-11|Cot1-546 (ug)]] | |||
|- style="font-size:12pt" | |||
|style="font-weight:bold" height="15" valign="bottom" | A1 | |||
|style="font-weight:bold" align="center" valign="bottom" | 3.6 (30) | |||
| align="center" valign="bottom" | X | |||
|style="font-weight:bold" align="center" valign="bottom" | 2.9 (30) | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
|- style="background-color:#D9D9D9;font-size:12pt" | |||
|style="font-weight:bold" height="15" valign="bottom" | A2 | |||
| align="center" valign="bottom" | X | |||
|style="font-weight:bold" align="center" valign="bottom" | 2.9 (30) | |||
| align="center" valign="bottom" | X | |||
|style="font-weight:bold" align="center" valign="bottom" | 3.3 (30) | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
|- style="font-size:12pt" | |||
|style="font-weight:bold" height="15" valign="bottom" | B1 | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
|style="font-weight:bold" align="center" valign="bottom" | 3.7 (30) | |||
| align="center" valign="bottom" | X | |||
|style="font-weight:bold" align="center" valign="bottom" | 7.9 (0.4) | |||
|- style="background-color:#D9D9D9;font-size:12pt" | |||
|style="font-weight:bold" height="15" valign="bottom" | B2 | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
| align="center" valign="bottom" | X | |||
|style="font-weight:bold" align="center" valign="bottom" | 21.4 (30) | |||
|style="font-weight:bold" align="center" valign="bottom" | 7.9 (0.4) | |||
|} | |||
#Allow stored slides to warm to room temperature | #Allow stored slides to warm to room temperature | ||
Line 19: | Line 71: | ||
#Denature for 2.5 minutes at 92C; keep humid | #Denature for 2.5 minutes at 92C; keep humid | ||
#Transfer slides to a humidified chamber and hybridize overnight at 37C or 42C | #Transfer slides to a humidified chamber and hybridize overnight at 37C or 42C | ||
[[Category:HRCP]] [[Category:Hybridization]] [[Category:20140919]] |
Latest revision as of 23:52, 23 September 2014
FISH[edit]
Today I'll perform the FISH experiments on the slides I prepared Wednesday
Hybridization[edit]
Sample Matrix
Sample | GAD1-488 (pmol) | FOXP2-546 (pmol) | RBFOX3-594 (pmol) | CUX2-647 (pmol) | DMD-B-488 (pmol) | PPRL-A2-488 (pmol) | Cot1-546 (ug) |
A1 | 3.6 (30) | X | 2.9 (30) | X | X | X | X |
A2 | X | 2.9 (30) | X | 3.3 (30) | X | X | X |
B1 | X | X | X | X | 3.7 (30) | X | 7.9 (0.4) |
B2 | X | X | X | X | X | 21.4 (30) | 7.9 (0.4) |
- Allow stored slides to warm to room temperature
- Incubate slides in 2X SSCT +50% (v/v) formamide for 2.5 minutes at 92 C in a pre-warmed coplin jar
- Incubate in coplin jar containing 2X SSCT + 50% formamide at 60C for 20 minutes
- Remove slides and allow to cool to room temperature
- Add 25 uL hybridization cocktail to a 22x22 #1.5 coverslip
- Invert slides on to cocktail-covered coverslips; seal with rubber cement
- Allow the rubber cement to air-dry for 5 minutes at room temperature
- Denature for 2.5 minutes at 92C; keep humid
- Transfer slides to a humidified chamber and hybridize overnight at 37C or 42C