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Protocols & Weblinks

Selection of protocols we frequently use:


ChIP, RT-qPCR and RNA-seq

  • ChIP
  • RT-qPCR
  • cDNA library for RNAseq
  • RNAseq analysis with Galaxy

Live-cell imaging

  • Pombe microscopy (agarose pads)
  • Pombe microscopy (sample preparation,
    imaging, analysis)

Ubiquitin pulldown and protein extracts

  • Ni-Ub pulldown under denat. conditions
  • Protein extraction w/ NaOH/TCA
  • Preparation of chromatin-enriched fraction
  • NuPage protein gel casting protocol

Large-scale protein purification of chromatin-associated proteins

  • Large-scale IP for MS
  • Small-scale IP
  • co-IP

Cloning & PCR

  • Sequence- & ligation-independent cloning (SLIC)
  • Primer design for KOs, tagging and qPCR
  • Perl script for KO primers (coming soon)

Yeast methods

  • Yeast transformation
  • Pombe transformation
  • Random spore analysis
  • Colony PCR w zymolyase
  • Yeast DNA prep (‘smash & grab’)
  • large-scale genomic DNA preparation
  • Yeast plating assays with pin-frogger (serial dilutions)

SGA with Singer RoToR HDA

  • RoToR SGA protocol for S. pombe
  • Plates & pads cleaning

Useful links:

  • Dept. Physiological Chemistry
  • Biomedical Center (BMC)
  • SFB 1064 Chromatin Dynamics
  • EpiGeneSys
  • Pombase
  • PombeNet (Forsburg lab)
  • Bahlerlab resources
  • Pubmed
  • Blast
  • Primer-3
  • Bionumbers
  • Venn diagrams
  • Stackoverflow
  • Github
  • Websupplement to Bioneer deletion library

 
Braun Lab • Biomedical Center • LMU Munich • Großhaderner Str. 9 • 82152 Martinsried • Germany • BraunLab.de

 
Last update: 13/01/2021 Disclaimer