siRNA EHF

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	siRNA concentration-30nM Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio). Incubate for 5 minutes at room temperature and add to cells. Incubate cells for 2 days at 37°C.

Protocol tips
siRNA concentration-30nM Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio). Incubate for 5 minutes at room temperature and add to cells. Incubate cells for 2 days at 37°C.

Publication protocol

Calu-3 cells were treated with 30nM Silencer Select negative control siRNA #2 or EHF siRNA (s25399, both from Ambion) using RNAiMax transfection reagent (LT). 48 h post-transfection, RNA was isolated from five samples per treatment using Trizol (LT). RNA-seq libraries were prepared using the TruSeq RNA Sample Preparation Kit v2 per the manufacturer's Low-Throughput protocol (Illumina). The libraries were sequenced on Illumina HiSeq2500 machines. Data were analysed using TopHat and Cufflinks (30). qRT-PCR was performed with TaqMan Reverse Transcription reagents kit (LT), primer pairs specific for each target (Supplementary Table S2) and SYBR green. All data are deposited at GEO (http://www.ncbi.nlm.nih.gov/geo/).

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing siRNA / miRNA gene silencing Human - Calu-3 EHF using siRNA EHF from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for siRNA EHF below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing siRNA / miRNA gene silencing Human - Calu-3 EHF using siRNA EHF from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.