Protocols

Many of these protocols have been updated and new ones were added in May 2018. Over the next year there will be further additions particularly for replicative ageing cell work and library preparation.

Basic protocols for new lab workers:
 
Making solutions
Handling enzymes
Agarose gels for DNA - updated May 2018
Basic rules for RNA work
 
Standard molecular biology protocols:
 
Cloning - updated May 2018
PCR
RT-PCR
PicoGreen quantification
qPCR - NEW May 2018
 
Blotting and probing protocols:
 
Genomic Southern Blot
Glyoxal gel
Denaturing PAGE
In vitro transcription - updated May 2018
Ladder labelling
Probing blots - updated May 2018
Western blotting - updated May 2018
siRNA blotting

Library preparation protocols:

mRNAseq library preparation - NEW May 2018
KAPA quantification - NEW May 2018

Yeast work:
 
Yeast culture - updated May 2018
Yeast genetic engineering
Yeast Transformation - NEW May 2018
Harvesting yeast - updated May 2018
Synchronised meiosis
BY4741 sporulation
Yeast Colony PCR - updated May 2018
 
Yeast DNA isolation
Yeast PFGE - updated May 2018
 
Which RNA prep do I use? Probably hot phenol, but GTC can be better for ribosome analysis. The GTC miniprep is the fastest and most consistent if you need to prep many samples and aren't too fussy about perfect quantification. With few samples, consider mirVANA kits – these produce beautiful RNA but are expensive
 
Yeast hot phenol RNA prep
Yeast hot phenol RNA miniprep
Yeast GTC RNA prep (Tollervey lab standard prep)
Yeast GTC RNA miniprep - updated May 2018
Yeast mirVANA total RNA prep - NEW May 2018
 
Yeast propidium iodide staining for FACS
ChIP for yeast
Yeast immunofluorescence
 
Yeast protein isolation - updated May 2018
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.