Daniel:Notebook/HiResChrPaint/2014-9-19: Difference between revisions

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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]

Back to Calendar

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)
  1. Allow stored slides to warm to room temperature
  2. Incubate slides in 2X SSCT +50% (v/v) formamide for 2.5 minutes at 92 C in a pre-warmed coplin jar
  3. Incubate in coplin jar containing 2X SSCT + 50% formamide at 60C for 20 minutes
  4. Remove slides and allow to cool to room temperature
  5. Add 25 uL hybridization cocktail to a 22x22 #1.5 coverslip
  6. Invert slides on to cocktail-covered coverslips; seal with rubber cement
  7. Allow the rubber cement to air-dry for 5 minutes at room temperature
  8. Denature for 2.5 minutes at 92C; keep humid
  9. Transfer slides to a humidified chamber and hybridize overnight at 37C or 42C