Highly sensitive biotinylated RNA standard suitable for bacterial small RNA (sRNA) research.
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.
Part of The Investigator's Annexe program.
|Product Type:||Buffer or Chemical|
|Name:||Biotinylated sRNA ladder|
|Buffer:||Liquid in TE buffer|
|Marker Size:||100, 200, 300, 400, 500, 750, 1000 bases|
|Comments:||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.|
Absorbance of sRNA Ladder
The purity and concentration of the standard was deteremined using a Nanodrop ND-1000. The volume of the stock is 100 uL the RNA is estimated at 1200 ng/uL with a 260nm/280nm ratio of 1.93 and a 260nm/230nm ratio of 2.26. The total amount of RNA was 120 ug.
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.
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.
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