In situ hybridisation and Immunohistochemistry

• Some general tips
• Probe preparation
  • David Wilkinson (RA and non RA)
  • KC Lab
• Section RNA ISH
  • KC Lab traditional RNA ISH protocol
  • Non-Radioactive ISH (DIG, Biotin, Fluorescein) (Qiling Xu)
  • Fluorescent ISH with oligo probes (from F.G lab Erasmus)
  • Non radioactive ISH protocols (sections, cells, wholemount)
• Whole mount RNA ISH
  • Koops
  • ICRF
• Wax embedding of embryos and tissue
• Y.ES Kit
• Antigen retrieval (Section IHC with notes Donald)
• Immunostaining of frozen sections of mouse embryos
• Section IHC (Woody, Chinese U)
• Whole mount immunostaining. (Dave Wilkinson '96)
• Slide Preparation
• X-Gal antibody
• Taking photos of sections
• Polyclonal type X antibody
• Peter Bruckners new type II ab (not tested)

Molecular biology

• DNA things
  • DNA preparation. All sorts of evils are covered in here, from Maxi and minipreps to isolation of frags from agarose gels
• Bug things
  • Shuttling plasmids
• RNA things
  • RNase protection assay
  • RT-PCR for very small quantities of cells
• PCR things
  • PCR (The Boerhinger-Mannheim guide to PCR)
  • Optimizing PCR
  • Purifying Taq from pTaq plasmid
  • Preparation of T-vectors
• Other molecular biology stuff
  • Home-made Chemiluminescent substrate (ECL)
  • Yeast Two Hybrid system
  • Preparation of nuclear extracts from mouse embryos
• Protein Work
  • Bio Rad semi dry blotter manual
    

Transgenic Production and maintenance

• X-Gal staining of embryos
• Alkaline Phosphatase staining of embryos
• Making mouse fibroblasts
• Nuclear Transfer from an Established Cell Line
• Mouse Embryo Freezing
• Preparation of M2 and M16

Misc

• Plasmid Shipment