m6a rna methylation quantification kit

RNA quantification Coloremetric

Experiment
RNA quantification Coloremetric
Product
m6a rna methylation quantification kit from Abcam
Manufacturer
Abcam

Protocol tips

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.

Publication protocol

Total m6A content was measured in 200-ng aliquots of total RNA extracted from MDA-MB-231 and MCF-7 subclones using an m6A RNA methylation quantification kit (cat. no. P-9005; Epigentek) according to the manufacturer’s instructions. To measure m6A+ NANOG mRNA levels, m6A immunoprecipitation was performed as described before (47). A 1-µg aliquot of m6A antibody was conjugated to protein A-agarose slurry (Millipore) overnight at 4 °C. A 100-μg aliquot of total RNA was incubated with the antibody in immunoprecipitation buffer (50 mM Tris⋅HCl, 750 mM NaCl, and 0.5% Igepal CA-630) supplemented with RNase inhibitor for 3 h at 4 °C, RNA was eluted from the beads by incubation in 300 μL of elution buffer (5 mM Tris⋅HCl, 1 mM EDTA, and 0.05% SDS) with 4.2 μL of proteinase K for 1.5 h at 50 °C, and m6A+ RNA was purified by phenol/chloroform extraction and analyzed by RT-qPCR. The nucleotide sequences of the primers used for detection for m6A+ NANOG mRNA were as follows: 5′-ATG CAA CCT GAA GAC GTG TG-3′ and 5′-GAG ATT GAC TGG ATG GGC AT-3′.

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing RNA quantification Coloremetric using m6a rna methylation quantification kit from Abcam.

Paper title
Hypoxia induces the breast cancer stem cell phenotype by HIF-dependent and ALKBH5-mediated mA-demethylation of NANOG mRNA
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Manufacturer protocol

Download the product protocol from Abcam for m6a rna methylation quantification kit below.

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