Biotinylated sRNA Ladder

Highly sensitive biotinylated RNA standard suitable for bacterial small RNA (sRNA) research.

Highlights:

  • Easily determine the size of bacterial sRNA without radiation
  • Highly sensitive probe (less than or ca. 12 ng/µL)
  • Suitable for sRNA bacterial and Archaea research and Northern Blot analysis

The discovery of bacterial small RNA (sRNA) is a major area of research in microbiology. Northern blotting remains the gold standard for sRNA identification since it confirms presence and size. To determine size using a northern blot it is necessary to run an RNA standard of known size. Typically, estimating the size of sRNA requires adding radioactive [U-32P]ATP to an RNA ladder using T4 polynucleotide kinase and imaging the resulting blot with film or a radioimager. The Biotinylated sRNA Ladder allows for a non-radioactive method of northern blotting using biotin linked probes and RNA ladders to identify and size small RNAs. The biotin linked probes work by adding a streptavidin-horsradish peroxidase conjugate to the blot then adding luminol and hydrogen peroxide to create chemiluminescence.

From the laboratory of Mary Ann Moran, PhD, Andrew S. Burns, PhD and Adam Rivers, PhD, University of Georgia.

The Investigator's Annexe Part of The Investigator's Annexe program.

Catalog Number Product Size AVAILABILITY Price Qty
EGA701
Biotinylated sRNA Ladder, 50uL (50 reactions)
50uL (50 rxns) In stock
Price: $188.00
Specifications
Name: Biotinylated sRNA ladder
Buffer: Liquid in TE buffer
Marker Size: 100, 200, 300, 400, 500, 750, 1000 bases
Concentration: 12 ng/uL
Comments: Use 1uL per lane
Storage: -80C
Shipped: Dry ice
Documentation

Suggested Protocol

For running of polyacrylamide gels, the Biotinylated sRNA Ladder (12ng/uL) should be mixed with equal volume of a RNA loading buffer. Then, use 1 µl of of the resulting solution per lane.

Data

Absorbance of sRNA Ladder

WB Data

The purity and concentration of the standard was deteremined using a Nanodrop ND-1000. The volume of the stock is 100 ?L the RNA is estimated at 1200 ng/?l with a 260nm/280nm ratio of 1.93 and a 260nm/230nm ratio of 2.26. The total amount of RNA was 120 ?g.

Northern Blot

WB Data

Northern blots of small RNAs from Rugerio Pomeroyi DSS-3. 1uL of 12 ng/uL standard or 20 ug of total bacterial RNA was added to each lane.

Provider
From the laboratory of Mary Ann Moran, PhD, Andrew S. Burns, PhD and Adam Rivers, PhD, University of Georgia.
References
  1. Burns AS, Bullock HA, Smith C, Huang Q, Whitman WB, Moran MA. Small RNAsexpressed during dimethylsulfoniopropionate degradation by a model marinebacterium. Environ Microbiol Rep. 2016 Jun 23. View Article
  2. Stacey SD, Pritchett CL. Pseudomonas aeruginosa AlgU Contributes toPosttranscriptional Activity by Increasing rsmA Expression in a mucA22 Strain. J Bacteriol. 2016 Jun 13;198(13):1812-26. View Article
  3. Rivers, A. R., Burns, A. S., Chan, L.-K., and Moran, M. A. (2016). Experimental Identification of Small Non-Coding RNAs in the Model Marine Bacterium Ruegeria pomeroyi DSS-3. Front. Microbiol. 7. doi:10.3389/fmicb.2016.00380.
  4. Gottesman S, Storz G. Bacterial small RNA regulators: versatile roles and rapidly evolving variations. Cold Spring Harb Perspect Biol. 2011 Dec 1;3(12).
  5. Locati MD, Pagano JF, Ensink WA, van Olst M, van Leeuwen S, Nehrdich U, Zhu K, Spaink HP, Girard G, Rauwerda H, Jonker MJ, Dekker RJ, Breit TM. Linking Maternal and Somatic 5S rRNA types with Different Sequence-Specific Non-LTR Retrotransposons. RNA. 2016 Dec 21. pii: rna.059642.116. View Article

If you publish research with this product, please let us know so we can cite your paper.

 
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