Protocols

Basic protocols for new lab workers:
 
Making solutions
Handling enzymes
Agarose gels for DNA
Basic rules for RNA work
 
Standard molecular biology protocols:
 
Cloning
PCR
RT-PCR
PicoGreen quantification
 
Blotting and probing protocols:
 
Genomic Southern Blot
Glyoxal gel
Denaturing PAGE
In vitro transcription
Ladder labelling
Probing blots
Western blotting

Yeast work:
 
Yeast culture
Yeast genetic engineering
Harvesting yeast
Synchronised meiosis
BY4741 sporulation
Yeast Colony PCR
 
Yeast DNA isolation
Yeast PFGE
 
Which RNA prep do I use? Probably hot phenol, but GTC can be better for ribosome analysis. With few samples, consider mirVANA kits – these produce beautiful RNA but are expensive – see siRNA blotting protocol for adapting them to yeast.
 
Yeast hot phenol RNA prep
Yeast hot phenol RNA miniprep
Yeast GTC RNA prep (Tollervey lab standard prep)
Yeast GTC RNA miniprep
siRNA blotting
 
Yeast propidium iodide staining for FACS
ChIP for yeast
Yeast immunofluorescence
 
Yeast protein isolation
TAP purification


These are our in-house protocols - they are provided as is and we accept no responsibility for their use or mis-use. Please observe your organisation's health and safety policy while performing these procedures.