RNA quantification Coloremetric

RNA quantification for appropriate concentration and quality (260/280 ratio) is an important step before downstream analysis (including sequencing, RT-qPCR, etc.). Having insufficient RNA quantities or a high salt or phenol in the RNA product can lead to variable or irreproducible downstream results. The various methods used for RNA quantification include: 1. UV spectrophotometric (challenges include: low sensitivity, cannot distinguish between nucleic acid species), 2. Fluorescence-based (challenges include: requires standards, cannot measure amplifiability, not sequence-specific), and 3. RT-PCR (challenges include: requires standards, time-intensive, costly). In order to overcome these challenges, and also to ensure the proper quantification and quality control for RNA product, it is important to use at least two or more methods in order to discard any inconsistencies. Using standards for calibrations increases the sensitivity range for RNA detention (fluorescence- and RT-PCR-based methods). When using RT- PCR, it is important to choose correct primers, aligning to the desired site on the template and of appropriate product length, along with positive, negative and loading controls. It is also important to have at least two primer pairs in order to confirm results.

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Found 7 matching solutions for this experiment

Protocol tips
A549 and PC9 cells transfected with or without siRNA against circABCB10 and a FOXR2 overexpression vector were seeded into 96‐well plates at 5 × 103 cells/well and incubated for increasing durations (0, 12, 24, 48, and 72 hours) before adding 10 μL of Cell Counting Kit‐8 (CCK8) solution per well for 1 hour. Absorbance (450 nm) was measured using a Synergy microplate reader (BioTek, Winooski, VT).
Upstream tips
This kit can detect 10^10 viral particles/mL from 100 µL of lentiviral supernatant.

This kit measure both the FIV and HIV virus particles
Protocol tips
Add QuickTiter Solution A to the
sample and incubate at 37ºC for 30 minutes
Downstream tips
Read the plate with a
fluorescence plate reader using a 480/520 nm filter set.
Upstream tips
This kit detects N
6-methyladenosine (m
6A) RNA methylation status directly using total RNA isolated from any species
such as mammals, plants, fungi, bacteria, and viruses.
Downstream tips
Read on a microplate reader at 450 nm within 2 to 15 min.
Upstream tips
This kit enables quantitation of as little as 1 ng/mL.
Protocol tips
Add 1.0 mL of the aqueous working solution of the Quant-iT™ Ribo Green® reagent and incubate for 2 to 5 minutes at room temperature, protected from light..
Qubit RNA HS Assay Kit

Thermo Fisher Scientific

Upstream tips
The Qubit® assays are designed to be performed at room temperature, as temperature fluctuations can influence the accuracy of the assay.
Protocol tips
Dilute Qubit® RNA HS Reagent 1:200 in Qubit® RNA HS Buffer.
Qubit RNA BR Assay Kit

Thermo Fisher Scientific

Upstream tips
This kit is easy to use and
accurate providing an assay range of 20–1000 ng.

It is intended for total RNA,
rRNA, or large mRNA
Protocol tips
Dilute the Qubit RNA BR Reagent 1:200 in Qubit RNA BR Buffer
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.
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