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Yeast in situ Hybridization
Published: Sep 20, 2011 DOI: 10.21769/BioProtoc.134 Views: 11783
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Abstract
This protocol describes the procedure to label a certain mRNA species in yeast cells with fluorescence probes and prepare slides for visualization under microscope. Probe design/synthesis and microscopy operation are not covered.
Materials and Reagents
- HCl
- Poly L-Iysine
- Formaldehyde
- β-mercaptoethanol
- Oxalyticase
- SDS
- 70% ethanol
- Formamide
- 20x SSC
- NaH2PO4 buffer
- Triton X-100
- 1x PBS-DEPC
- DAPI
- 40% formamide/2x SSC (see Recipes)
- 80% formamide (see Recipes)
- Hybridization solution (see Recipes)
Equipment
- Incubator
- Centrifuges
- Glass beaker
- Autoclaved glass cylinder
- Baked glass pipette
- Hotplate
- Fume hood
- Petri dish
- Foil
- 50 ml conical tubes
- Parafilm
- Light microscope
Procedure
Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.
Category
Microbiology > Microbial biochemistry > RNA
Microbiology > Microbial genetics > RNA
Molecular Biology > RNA > RNA labeling
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