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Tools and Skills

Cell Biology 

  • Clonogenic cell survival assays
  • Viability assays (SRB, CellTiterGlo, MTT, WST)
  • Analysis of DNA end resection by flow cytometry
  • Analysis of cell cycle progression and checkpoints

Molecular Biology 

  • Q-PCR 
  • site directed mutagenesis 
  • inducible shRNA expression 
  • (FP/FLAG/HA/Myc/His)-tagged proteins
  • CRISPR/Cas9 technology for genome editing and DSB induction

Cell Imaging 

  • Biphoton laser microirradiation (DNA damage induction, live cell imaging for protein   recruitment and protein mobility (FRAP)) 
  • Reporter systems for DNA double-strand break repair (HR, End-Joining, SSA) in living cells
  • High-resolution imaging of DNA repair complexes (Ku & DNA-PKcs foci)
  • Super-resolution imaging of DNA repair complexes (3D-SIM & STORM)
  • Imaging of clickable small molecules

Biochemistry 

  • Electrophoretic Mobility Shift Assay
  • DNA-PK kinase assay 
  • DNA end synapsis assay
  • In vitro end-joining reactions on linear plasmids
  • Protein production (bacteria and insect cells) and purification
  • Identification of drug targets by chemoproteomics

 

Telomere Analysis 

  • Telomeric FISH 
  • Shelterin Immunofluorescence 
  • IF-FISH 
  • Telomeric restriction fragments (TRF) 
  • STELA 
  • Telomerase assay