, For more technical information about the parameters, download the supplier's pipette cookbook

, hold the pipette at each extremity and heat the thinnest part of it on the flame for few seconds, turning continuously the pipette until the thin part becomes red. When it starts to melt, remove quickly from the flame and pull constantly until ductility is lost. Then, cut the thinner Pasteur pipette at an appropriate distance to fill the micropipette

, To improve fly survival, it is best not to leave flies more than half an hour on the CO 2anaesthetizing pad. We suggest injecting flies in small groups

, Injection can be performed intra-abdominally (6), but in our hands lethality after injection is higher

, Melanization spots appear a few minutes after injury, but for convenience and to prevent repeated anaesthesia at the same day, you should check the day after. For an overview of melanization, see ref

A. Fire, S. Xu, M. K. Montgomery, S. A. Kostas, S. E. Driver et al., Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans, Nature, vol.391, pp.806-811, 1998.

P. M. Waterhouse, M. W. Graham, W. , and M. B. , Virus resistance and gene silencing in plants can be induced by simultaneous expression of sense and antisense RNA, Proc Natl Acad Sci U S A, vol.95, pp.13959-13964, 1998.

J. R. Kennerdell and R. W. Carthew, Use of dsRNA-mediated genetic interference to demonstrate that frizzled and frizzled 2 act in the wingless pathway, Cell, vol.95, pp.1017-1026, 1998.

H. Siomi and M. C. Siomi, On the road to reading the RNA-interference code, Nature, vol.457, pp.396-404, 2009.

S. C. Miller, S. J. Brown, and Y. Tomoyasu, Larval RNAi in Drosophila?, Dev Genes Evol, vol.218, pp.505-510, 2008.

S. Dzitoyeva, N. Dimitrijevic, and H. Manev, Intra-abdominal injection of doublestranded RNA into anesthetized adult Drosophila triggers RNA interference in the central nervous system, Mol Psychiatry, vol.6, pp.665-670, 2001.

A. Goto, S. Blandin, J. Royet, J. M. Reichhart, and E. A. Levashina, Silencing of Toll pathway components by direct injection of double-stranded RNA into Drosophila adult flies, Nucleic Acids Res, vol.31, pp.6619-6623, 2003.
URL : https://hal.archives-ouvertes.fr/hal-00311256

S. Dzitoyeva, N. Dimitrijevic, and H. Manev, Gamma-aminobutyric acid B receptor 1 mediates behavior-impairing actions of alcohol in Drosophila: adult RNA interference and pharmacological evidence, Proc Natl Acad Sci U S A, vol.100, pp.5485-5490, 2003.

M. C. Saleh, R. P. Van-rij, A. Hekele, A. Gillis, E. Foley et al., The endocytic pathway mediates cell entry of dsRNA to induce RNAi silencing, Nat Cell Biol, vol.8, pp.793-802, 2006.

J. Ulvila, M. Parikka, A. Kleino, R. Sormunen, R. A. Ezekowitz et al., Double-stranded RNA is internalized by scavenger receptor-mediated endocytosis in Drosophila S2 cells, J Biol Chem, vol.281, pp.14370-14375, 2006.

J. S. Whangbo and C. P. Hunter, Environmental RNA interference, Trends Genet, vol.24, pp.297-305, 2008.

X. H. Wang, R. Aliyari, W. X. Li, H. W. Li, K. Kim et al., RNA interference directs innate immunity against viruses in adult Drosophila, Science, vol.312, pp.452-454, 2006.

D. Galiana-arnoux, C. Dostert, A. Schneemann, J. A. Hoffmann, and J. L. Imler, Essential function in vivo for Dicer-2 in host defense against RNA viruses in drosophila, Nat Immunol, vol.7, pp.590-597, 2006.

R. P. Van-rij, M. C. Saleh, B. Berry, C. Foo, A. Houk et al., The RNA silencing endonuclease Argonaute 2 mediates specific antiviral immunity in Drosophila melanogaster, Genes Dev, vol.20, pp.2985-2995, 2006.
URL : https://hal.archives-ouvertes.fr/pasteur-01954329

M. C. Saleh, M. Tassetto, R. P. Van-rij, B. Goic, V. Gausson et al., Antiviral immunity in Drosophila requires systemic RNA interference spread, Nature, vol.458, pp.346-350, 2009.
DOI : 10.1038/nature07712
URL : https://hal.archives-ouvertes.fr/pasteur-00460853

I. Sanchez-vargas, J. C. Scott, B. K. Poole-smith, A. W. Franz, V. Barbosa-solomieu et al., Dengue virus type 2 infections of Aedes aegypti are modulated by the mosquito's RNA interference pathway, PLoS Pathog, vol.5, p.1000299, 2009.

H. Tang, Regulation and function of the melanization reaction in Drosophila, Fly (Austin), vol.3, pp.105-111, 2009.