Selection of protocols & methods

ChIP, RT-qPCR and RNA-seq

Live-cell imaging

Pombe microscopy (sample preparation, imaging, analysis) (coming soon)

Ubiquitin pulldown and protein extracts

Large-scale protein purification of chromatin-associated proteins

Cloning & PCR

Primer design for KOs, tagging and qPCR overview on primer design for different purposes
Perl script for KO primer design incl. README.md how to run script and info about primer design

Yeast methods

Yeast transformation S. cerevisiae transformation protocol by the Madhani lab (aka Yeast 2000)
Pombe transformation protocol adapted from the Krogan lab
Random spore analysis adapted from Susan L. Forsburg and Nicholas Rhind (Yeast 2006)
Yeast DNA prep (‘smash & grab’) phenol-based protocol, optional EtOH precipitation; works for Sc and Sp
Large-scale genomic DNA preparation phenol-chloroform based protocol, for high-yield and quality genomic DNA
Colony PCR w zymolyase fast & simple protocol for PCR-based genomic DNA analysis (e.g. deletions, epitope-tags)
Telo PCR for telomere-length measurements, adapted from the Azzalin lab
Yeast plating assays serial dilution-based growth assays using 48- or 96-pin tools; fast and easily scalable

SGA with Singer RoToR HDA