Techniques with DNA

• DNA preparation
  - isolation of plasmid DNA from yeast and bacteria
  - isolation of genomic DNA from yeast or murine cells
  - isolation of phage DNA

• DNA manipulation
  - cloning
  - restriction digest
  - restriction mapping of genomic DNA

• DNA analyses
  - by gel electrophoresis
  - by Southern blotting using plasmid or genomic DNA
  - by South-Western protocol

•  DNA sequencing
  - Maxam Gilbert sequencing
  - Sanger dideoxy sequencing

•  Probe preparation
  - end-labelling
  - random priming
  - fill-in labelling
  - nick-translation

• Library screening
  - direct screening (southern method)
  - expression library screening (south-western protocol)

Protein Biochemistry

•  Protein/DNA interaction in vitro
  - by DNAse1 footprinting
  - by hydroxyl radical - footprinting
  - by Electrophoretic mobility shift assay
  - by South-Western blotting
  - by methylation interference
  - by ethylation interference

• Protein/DNA interactions in vivo
  - using One-Hybrid system

• Protein/Protein interaction in vitro
  - by immunoprecipitation
  - by GST-pull-down

•  Protein/Protein interaction in vivo
  - using Two-hybrid system

•  Recombinant protein expression/isolation
  - of fusions with GST, MBP, His-tag
  - familiar with production of small scale protein expression systems in baculovirus

• Recombinant protein analyses
  - by SDS-PAGE

• see the COMPUTER SKILLS section below

Techniques with RNA

• Isolation of total RNA or mRNA

• cDNA synthesis

• 5’-RACE (rapid amplification of cDNA ends)

• 3’-RACE

Cell Biology

• Cell culture
  - in mammalian (Hep3B, HepG2, COS1) and avian (LMH/2A) cell lines

• Experience working with both S. cerevisiae and S. pombe

PCR

• RT-PCR

• PCR from plasmid, genomic samples

• PCR in mutagenesis
  - by site-directed mutagenesis
  - by recombinant mutagenesis

Immunochemistry

• Western blotting

• Generation of polyclonal antibodies and subsequent purification

• Immunocytochemistry in transfected cells