The RNA Ladder, Low Range is a mixture of seven chromatography-purified single-stranded RNA transcripts. Their lengths are (in bases ): 1000, 800, 600, 400, 300, 200 and 100. The ladder is designed for qualitative and quantitative analysis of RNA on agarose gels stained with ethidium bromide. The ladder is free of degraded RNA and NTP’s. Therefore, spectrophotometric measurements provide accurate values of RNA concentration in each ladder band (see photos for quantitative data). Due to this feature, the RNA ladder could be used for approximate RNA quantification on gels. RNA Ladder, Low Range is produced from templates that contain a fragment of the pTZ19R polylinker and Lambda phage fragments.||Features:|Sharp bands of uniform intensity|Easy-to-remember band sizes and quantities|Approximate RNA quantification by predetermined concentration of ladder bands|Supplied with 2X RNA Loading Dye|Stable for 6 months at -20°C||Applications:|RNA sizing and quantification on native or denaturing gels|Northern blotting||Usage:|The RNA Ladder is recommended for EP in the following: native 2% agarose with TAE buffer, denaturing formaldehyde agarose with MOPS buffer, denaturing glyoxal/DMSO agarose with sodium phosphate buffer and denaturing polyacrylamide gel electrophoresis in TBE buffer.||Quality Control Assay Data:|Analysis of 2ul of RNA Ladder, Low Range, on a polyacrylamide gel and native agarose gel with ethidium bromide staining generates a clear seven-band pattern.|Caution: This product is extremely sensitive to degradation by ribonucleases. The use of diethyl pyrocarbonate-treated solutions and protective gloves is recommended.||Storage and Stability:|Aliquot to avoid repeated freezing and thawing and store at -70°C. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. ||Protocol:|1. Thaw the ladder on ice.|2. Mix the contents well before use by pipetting or gentle vortexing as concentration gradients may form in frozen products over time. |3. Use a 0.25ul aliquot of the ladder per 1mm of the gel lane width. |4. Prepare the following for 8mm width of gel lane:| 2ul of R2033-12A1 Loading Dye| 2ul of R2033-12 Ladder|5. Vortex briefly and spin down. |6. Heat at 70°C for 10 min. Chill quickly on ice and load on gel.||Protocol Notes:|he Loading Dye allows for RNA visualization without additional staining of denaturing agarose gels. If RNA fragments are separated on native agarose gels, additional staining with ethidium bromide is recommended.|• When visualizing a gel under UV light, an additional dark zone of ethidium bromide can sometimes be observed. However, this has no influence on the quality of RNA separation.|• Avoid long exposure to the UV light, as this may cause RNA degradation.|• For Northern blots, perform EP in denaturing formaldehyde agarose with MOPS buffer. A 2ul aliquot of the RNA Ladder is well visible after being transferred on Hybond-N+ membrane from 2% formaldehyde gel, whereas the same amount of RNA ladder is less visible when transferred from 2% native agarose.|• The ethidium bromide present in the sample and/or in the gel does not interfere neither with the RNA transfer onto the membrane, nor with RNA hybridization with the probe.