Institut Jacques Monod

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Protocoles


Biologie moléculaire

preparation of genomic DNA:  
quick protocol for single flies (very fast, good to amplify fragments smaller than 1kb; this genomic DNA can only be kept for a few months at –20C. Use another protocol if you want to amplify larger fragments or keep the DNA longer)
quick protocol for fly wings (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 using DNAzol DNAzol sheet 1 DNAzol sheet 2

extraction of RNA: using Promega kit

preparation of cDNA: using Ambion kit (did not work well), other protocol using 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, antibody staining, 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  for details about in 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 new protocol    my post-doc protocol      my PhD protocol
non fluorescent in situ staining of embryos: old protocol  protocol in C. D-V. lab
protocols from Leslie Vosshall


Fly food
Banana-opuntia fly food recipe



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



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


Yeast protocols
yeast media recipes html PDF
geneticin
my protocols

primers erg2 mutants
yeast deletion strains


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


Manuals

RNAlater
pGEM-T (PDF)
low mass ladder

plasmid maps:
  pBluescript II KS-
  pBluescript II SK-
  pBluescript II SK+
  pcaSpeR-hs