Northern Blot Analysis of RNA

Total RNA was extracted and prepared as described previously^{58 (link)}. For Northern blot analysis, RNA samples were separated on 6% polyacrylamide/7 M urea gels and transferred to Hybond-XL membranes (GE Healthcare). Membranes were hybridized in Roti-Hybri-Quick buffer (Roth) at 42 °C with [³²P] end-labeled DNA oligonucleotides. Oligonucleotides used for probing are listed in Supplementary Table S4. Membranes were washed in three subsequent steps with SSC (5x, 1x, 0.5x)/0.1% SDS wash buffer. Signals were visualized on a Amersham Typhoon phosphorimager (GE Healthcare) and quantified with GelQuant software (BiochemLabSolutions).

Free full text: Click here

Huber M., Lippegaus A., Melamed S., Siemers M., Wucher B.R., Hoyos M., Nadell C., Storz G, & Papenfort K. (2022). An RNA sponge controls quorum sensing dynamics and biofilm formation in Vibrio cholerae. Nature Communications, 13, 7585.

Publication 2022

Hybond-XL membranes Roti-Hybri-Quick buffer Amersham Typhoon phosphorimager

Buffer Membranes Northern blot analysis Oligonucleotides Polyacrylamide gels Typhoon Urea

Corresponding Organization :

Other organizations : Friedrich Schiller University Jena, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Dartmouth College

Top 5 similar protocols

Variable analysis

independent variables

Preparation of total RNA samples

dependent variables

Expression levels of target RNAs as measured by Northern blot analysis

control variables

Conditions for polyacrylamide/urea gel electrophoresis
Conditions for RNA transfer to Hybond-XL membranes
Conditions for hybridization of radioactively labeled DNA oligonucleotide probes
Conditions for membrane washing steps
Conditions for signal visualization and quantification

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!