Hs_PLK1_7 FlexiTube siRNA

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	siRNA-Hs_PLK1_7: CGCGGGCAAGATTGTGCCTAA Final concentration of 15 nM each, or as a pool of three siRNAs (denoted as siHES6) 10 nM each. Incubate the FuGENE 6 Transfection Reagent/medium mixture for 5 minutes at room temperature  Incubate transfection mixture for 15 min at room temperature and add to cells and shake for 10-30 seconds.  Incubate cells for 24-48 h.

Protocol tips
siRNA-Hs_PLK1_7: CGCGGGCAAGATTGTGCCTAA Final concentration of 15 nM each, or as a pool of three siRNAs (denoted as siHES6) 10 nM each. Incubate the FuGENE 6 Transfection Reagent/medium mixture for 5 minutes at room temperature  Incubate transfection mixture for 15 min at room temperature and add to cells and shake for 10-30 seconds.  Incubate cells for 24-48 h.

Publication protocol

Cell lines A172 and U87MG were obtained from ECACC (Salisbury, UK), LN405 from the DSMZ (Braunschweig, Germany) and SVG p12 from ATCC (Manassas, VA, USA). Cells were cultured in medium conditions recommended by the providers for less than 4 months before use in these experiments. Sequences for three siRNAs for HES6 and a control siRNA for PLK1 are described in supplementary materials. All siRNAs were purchased from Qiagen (Hilden, Germany). Additional control siRNAs were AllStars Hs Cell Death Control siRNA (Qiagen) and Negative Control siRNA (Qiagen). The siRNAs were used at a final concentration of 15 nM each, or as a pool of three siRNAs (denoted as siHES6) 10 nM each, thus giving a final concentration of 30 nM. LN405 cells were used for creating stable cell lines by transfecting pEYFP-C1-mock and pEYFP-C1-HES6 with Fugene6 (Roche Applied Science, Indianapolis, IN, USA) and selecting the positive cells with 600 μg/ml and maintained with 400 μg/ml of G418 (Sigma-Aldrich, St Louis, MO, USA)

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Manufacturer protocol

Download the product protocol from Qiagen for Hs_PLK1_7 FlexiTube siRNA below.

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