Daniel:Notebook/HiResChrPaint/2014-11-4: Difference between revisions
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>Djacobse (Created page with "=FISH= Back to Calendar ==Hybridization== '''Sample Matrix''' {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/...") |
>Djacobse |
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==Hybridization== | ==Hybridization== | ||
*Using the [[Daniel:Notebook/HiResChrPaint/2014-10-2|Beliveau SSC Buffer]] | |||
'''Sample Matrix''' | '''Sample Matrix''' | ||
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#Allow the rubber cement to air-dry for 5 minutes at room temperature | #Allow the rubber cement to air-dry for 5 minutes at room temperature | ||
#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 | #Transfer slides to a humidified chamber and hybridize overnight at 37C | ||
[[Category:HRCP]] [[Category:Hybridization]] [[Category:20141104]] | [[Category:HRCP]] [[Category:Hybridization]] [[Category:20141104]] |
Latest revision as of 17:34, 5 November 2014
FISH[edit]
Hybridization[edit]
- Using the Beliveau SSC Buffer
Sample Matrix
Sample | uL DMD-A-488 (pmol) | uL Cot1-A-546 (ug) |
A | 3 (25.5) | 8 (0.4) |
B | 10 (85.5) | 8 (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