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Low Range ssRNA Ladder |
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Prices are in US dollars and valid only for US orders.
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Description:
The Low Range ssRNA Ladder is a set of 6 RNA molecules produced by in vitro transcription of a mixture of 6 linear DNA templates. The ladder sizes are: 1000, 500, 300, 150, 80 and 50 bases. The 300 base fragment is at double intensity to serve as a reference band. This ladder is suitable for use as an ssRNA size standard on denaturing or native agarose gels.
Usage Recommendation:
This marker was not designed for precise quantification of ssRNA mass.
Denaturing vs. Native Agarose Gels:
It is common practice to electrophorese RNA on a fully denaturing agarose gel, such as one containing formaldehyde (1). However, in many cases it is possible to run RNA on a native agarose gel and obtain suitable results. In fact, it has been demonstrated that treatment of RNA samples in a denaturing buffer maintains the RNA molecules in a denatured state, during electrophoresis, for at least 3 hours (2,3). The use of native agarose gels eliminates problems associated with toxic chemicals and the difficulties encountered when staining and blotting formaldehyde gels.
Figure 1: 1 µg of Low Range ssRNA Ladder was heated at 60°C for 5 minutes in 1X ssRNA Ladder Sample Buffer and visualized by ethidium bromide staining (2.0% TBE agarose gel).
Reagents Supplied:
ssRNA Ladder Loading Buffer (2X)
Concentration:
500 μg/ml
Storage Conditions
Storage Conditions:
20 mM KOAc
pH 4.5 @ 25°C
Storage Temperature:
-70°C
Notes
General notes:- Store at -70°C. For short term storage (< 1 week), ladder can be stored at -20°C.
- 2X ssRNA Ladder Sample Buffer no longer contains xylene cyanol ff.
Usage notes:- To avoid ribonuclease contamination: wear gloves, use RNase-free water for gels and buffers, wash equipment with detergent and rinse thoroughly with RNase-free water.
It is best to use freshly poured gels that are as thin as possible (i.e. 2-10 mm). Excessively long run times or high voltage can cause degradation of the bands on the gel. We recommend 4-8 volts/cm and running the bromophenol blue approximately 5 cm into the gel for good resolution.
Adding ethidium bromide to agarose gels and running buffer at a final concentration of 0.5 µg/ml will effectively stain the bands during electrophoresis.
FAQs
- How to prepare denatured RNA samples to be run on a native gel?
References
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., pp. 7.43?7.45. Cold Spring Harbor Laboratory Press.
- Liu, Y-C. and Chou, Y-C. (1990) Biotechniques, 9, 558.
- Sandra Cook, and Christina Marchetti, unpublished observations.
- Dong Ma, and John Greci, unpublished observations.
Reagents Sold Separately
ssRNA Ladder Loading Buffer
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