May 15, 2019
RNA concentration measurement and non-denaturing agarose gel electrophoresis
- Lisa-Maria Rosenthal1,
- Giang Tong1,
- Katharina Schmitt1
- 1Deutsches Herzzentrum Berlin, Berlin, Germany
External link: https://doi.org/10.1371/journal.pone.0226005
Protocol Citation: Lisa-Maria Rosenthal, Giang Tong, Katharina Schmitt 2019. RNA concentration measurement and non-denaturing agarose gel electrophoresis. protocols.io https://dx.doi.org/10.17504/protocols.io.2v5ge86
Manuscript citation:
Rosenthal L, Leithner C, Tong G, Streitberger KJ, Krech J, Storm C, Schmitt KRL (2019) RBM3 and CIRP expressions in targeted temperature management treated cardiac arrest patients—A prospective single center study. PLoS ONE 14(12): e0226005. doi: 10.1371/journal.pone.0226005
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 15, 2019
Last Modified: May 15, 2019
Protocol Integer ID: 23197
Abstract
RNA concentration measurement and non-denaturing agarose gel electrophoresis
Materials
MATERIALS
NanoDrop™ 2000c SpectrophotometerThermo Fisher ScientificCatalog #ND-2000C
• Allow RNA to thaw on ice
• Determine RNA concentration using a NanoDropTM 2000/2000c Spectrophotometers (Nucleic Acids – RNA mode, 2 µL sample size)
• Determine RNA purity from protein contamination by assuring 260/280 ratio is 2.0
• Determine RNA integrity and purity from genomic DNA contamination by electrophoresis. Load 400 ng RNA onto a 1% agarose gel with ethidium bromide (EtBr) in TBE buffer and run at 70 V until the dye line is approximately 75-80% of the way down the gel.
• Visualize and document using a gel UV imaging/documentation system