DRSC: Protocols

The DRSC periodically publishes papers describing our standard procedures for RNAi-based screening of Drosophila cells. These publications present detailed protocols in addition to tips on troubleshooting common problems, obtaining reagents and minimizing off-target effects.

Cell Screening Protocols from the DRSC:

Ramadan N, Flockhart I, Booker M, Perrimon N, Mathey-Prévôt B. Design and implementation of high-throughput RNAi screens in cultured Drosophila cells. Nat Protoc. 2007;2(9):2245-64.

Armknecht S, Boutros M, Kiger A, Nybakken K, Mathey-Prévôt B, Perrimon N. High-throughput RNA interference screens in Drosophila tissue culture cells. Methods Enzymol. 2005;392:55-73.

DRSC Publications on Off-Target Effects, False Positives and Reproducibility

Booker M, Samsonova AA, Kwon Y, Flockhart I, Mohr SE, Perrimon N. False negative rates in Drosophila cell-based RNAi screens: A case study. BMC Genomics 2011, 12:50doi:10.1186/1471-2164-12-50.

DasGupta R, Nybakken K, Booker M, Mathey-Prévôt B, Gonsalves F, Changkakoty B,Perrimon N. A case study of the reproducibility of transcriptional reporter cell-based RNAi screens in Drosophila. Genome Biol. 2007;8(9):R203.

Echeverri CJ, Beachy PA, Baum B, Boutros M, Buchholz F, Chanda SK, Downward J, Ellenberg J, Fraser AG, Hacohen N, Hahn WC, Jackson AL, Kiger A, Linsley PS, Lum L, Ma Y, Mathey-Prévôt B, Root DE, Sabatini DM, Taipale J, Perrimon N, Bernards R. Minimizing the risk of reporting false positives in large-scale RNAi screens. Nat Methods. 2006 Oct;3(10):777-9.

Kulkarni MM, Booker M, Silver SJ, Friedman A, Hong P, Perrimon N, Mathey-Prévôt B. Evidence of off-target effects associated with long dsRNAs in Drosophila melanogaster cell-based assays. Nat Methods. 2006 Oct;3(10):833-8.

Protocols for Cell-Based Screening in Drosophila

Cell Screening Protocols from the DRSC:

DRSC Publications on Off-Target Effects, False Positives and Reproducibility