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Protocols of V. Courtier-Orgogozo Lab

Molecular biology

preparation of genomic DNA:  
quick protocolfor singleflies (very fast, good to amplify fragments smaller than 1kb; this genomic DNA canonly be kept for a few months at –20C. Use another protocol if you want to amplify larger fragments or keep the DNA longer)
quick protocolfor flywings (great if you want to keep the flies alive!!!)
using QIAGEN DNeasy Tissue Kit (very good to amplify fragments that are 2-10kb long)
"blue book"protocol (good to amplify 2kb-PCR fragments, the result is a mix of genomic DNA+RNA (no RNAse step))
BDGP protocol
protocol usingDNAzol DNAzol sheet 1 DNAzol sheet 2

extraction of RNA: using Promega kit

preparation ofcDNA: using Ambion kit (did not work well), other protocolusing Promega kit (OK)
Inverse PCR:   BDGP protocol

PCR:  using ABgene PCR mix
        using Roche Expand High Fidelity
        using degenerate primers  (M. Koelle advices, David Towles protocol, HCGS protocol)
        PCR primer design  

general protocols:
   A. Moore lab
   C. D-V lab             

Staining protocols
muscle dye staining protocols

Antibody staining
antibody staining of embryos: my protocol
antibody staining of embryos with amplification: my protocol (PDF file)
antibody staining of larval gonads and other larval tissues: my protocol
antibody staining of pupal abdominal epidermis: N. Gompel's protocol (PDF file)
antibody staining of pupal abdominal muscles: my protocol
vizualisation of adult abdominal muscles (polarized light, antibodystaining, etc.): my protocol
antibody staining of adult ovaries: Horacio's protocol
                                              Emilie's protocol
list of antibodies

In situ staining
Check also  http://www-biology.ucsd.edu/~davek/index.html  fordetails aboutin situ hybridization
preparation of new in situ plasmids and probes: my protocol from RIKEN  my current protocol
fluorescent in situ+antibody staining of embryos: my newprotocol    my post-doc protocol      my PhDprotocol
non fluorescent in situ staining of embryos: old protocol  protocol inC. D-V. lab
protocols from Leslie Vosshall

Fly food
Banana-opuntia flyfood recipe

Fly organ measurements
my protocols for pupal cases, wings, palps, legs, dry mass
counting ovariolenumber

Fly genetics
Crosses and genetic markers for my D. simulans-D. sechellia QTL crosses

Yeast protocols
yeast media recipes html PDF
my protocols

primers erg2mutants
yeast deletion strains

Behavioral and other assays
resistance to octanoic acid
sperm motilityassay (PDF file with picture)


low massladder

plasmid maps:
  pBluescript IIKS-
  pBluescript IISK-
  pBluescript IISK+