Breaking Report: a lac operon system for expression in S2 cells

On my desk to read today? A report describing use of the lac operon system for inducible expression of a transgene in S2 cells.

Wakiyama et al. (2011) Inducible protein expression in Drosophila Schneider 2 cells using the lac operator-repressor system. Biotechnol. Lett. PMID: 21826399.

Breaking Report: Making dsRNA Vectors

Making dsRNA constructs in vectors? Hauge and colleagues report a one-tube, ligation-independent cloning method. If you're making your own constructs for in vitro or in vivo work, might be worth taking a look at their strategy.

Hauge et al. (2009) Single tube, high throughput cloning of inverted repeat constructs for double-stranded RNA expression. PLoS One. 4(9):e7205.

Transfection

The DRSC Protocols Pages describe our current methods for things like cell culture, transfection, dsRNA production and setting up a 384-well format screen. 

Today I'm thinking most about transfection. As with any protocol, I'm interested in two things. How to do it well. And how to do it well cheaply--particularly important when you're doing things at high-throughput scale as researchers do here for whole-genome screens.

Based on experience, we've been using Qiagen Effectene as a transfection reagent.  Some visiting screeners use a reagent from Invitrogen. And I recently got a protocol from the folks at Genesee Scientific for FectoFly (made in Europe by PolyPlus).

Have feedback? Experience? Favorites? We are interested to hear about it. So we can provide the best advice on the DRSC website and to visiting researchers.