RediPlate™ 96 RiboGreen™ RNA Quantitation Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Quantitation Range is 3-200 ng	Add 180 µL of the provided RediPlate TE buffer (Component B) to samples. Incubate samples for 10 minutes at room temperature, protected from light.

Upstream tips
Quantitation Range is 3-200 ng
Protocol tips
Add 180 µL of the provided RediPlate TE buffer (Component B) to samples. Incubate samples for 10 minutes at room temperature, protected from light.

Publication protocol

Briefly, DCs collected from the lower chamber were lysed, and total RNA was extracted using the RNAqueous Kit (Ambion, Paisley, United Kingdom) and Turbo DNase treatment (Ambion) according to the manufacturers’ recommendations. The quality of the RNA was verified on a Model 2100 Bioanalyzer (Agilent, Santa Clara, USA), and the RNA concentration was determined using a Rediplate 96 Ribogreen RNA Quantitation Kit (Gibco Invitrogen). Three samples were taken per treatment.

Real time RT-PCR analysis was carried out using a Human TLR Signaling Pathway PCR Array (SABiosciences Corporation, Frederick, Maryland, USA), which includes primer pairs specific for 20 selected genes related to TLR-mediated signaling pathways, as previously described by Bermudez-Brito et al. [14]. The expression level of each gene was analysed with RT2 Profiler PCR Array Data Analysis software (version 3.4; SABiosciences). The change in expression of each gene was reported as the fold change in expression (Fc). The results reflect logarithmic fold increase relative to the control samples.

Finally, IECs collected from the transwell membrane were lysed, and total RNA was extracted using the same method as described above. Real time RT-PCR analysis was carried out using a Human TLR Signaling Pathway PCR Array. The results are provided as [Additional files 1, 2, 3 and 4].

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for RediPlate™ 96 RiboGreen™ RNA Quantitation Kit below.

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