Product Name :Code No. :Range :
Size :

DynaMarker

RNA High
DM160
200-8,000 bases of RNA
50 g (56 l), 0.9 mg/ml

This product is research use only

Description :The DynaMarker RNA High consists of nine single-stranded RNAs, 200,500, 1,000, 1,500, 2,000, 3,000, 4,000, 5,000 and 8,000 bases, which
are synthesized by in vitro transcription. The DynaMarker RNA High issuitable for determinating size of single-stranded RNAs in denaturing
agarose gel electrophoresis. The concentration of each RNA(200-8,000 base) in the marker is approximately 0.1 g/l. It is usefulfor estimating of RNA amount. The DynaMarker RNA High can bevisualized by UV light after ethidium bromide staining or exposure tofilm with end labeling.

base
8,000
5,000
4,000
3,000
2,000
1,500
1,000
500

Storage buffer :10 mM Tris-HCl (pH 8.0) buffer containing 1 mM EDTA

200

Storage condition :Store at -80 oC. Repeated freeze/thaw cycles should be avoided.
DynaMarker

Quality Control : After 18 hr incubation of the DynaMarker RNA High at37 oC, no visible degradation of the marker is observed informaldehyde-agarose (1%) gel electrophoresis.

RNA High

Electrophoresis profile ofRNA High (0.9 g) on
formaldehyde-agarose (1%) gel

DynaMarker

Note :RNA is very sensitive to degradation by nucleases. To avoid damaging the DynaMarker RNA High, use extreme careduring manipulations to prevent nuclease contamination. Wear gloves and use clean apparatus. Glassware should
be pretreated with diethyl pyrocarbonate (DEPC). Nuclease-free disposable plasticware should be used.
Solutions and reagents to mix the marker should be high grade and nuclease-free. To use, thaw the DynaMarker RNAHigh on ice and keep it on ice while using.
Recommended usage :The DynaMarker RNA High is suitable for RNA size determining in denaturing agarose gel electrophoresis. For oneof example, DynaMarker RNA High can be run on denaturing agarose gel containing formaldehyde below.
Procedure
1. Agarose gel containing formaldehyde
Add 1 g of agarose to 85 ml of H2O in a flask, dissolve the agarose in a microwave. Add 10 ml of 10 × MOPS
buffer to the agarose solution, then allow it in a flask to cool to 55 oC. Add 5.4 ml of 37 % formaldehyde solution
to the agarose solution, mix them, quickly pour the agarose into a gel mold and set a comb in a fume hood.
Cover the gel with 1 × MOPS buffer until use.

1BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.6

Formaldehyde is supplied as a 37-40 % W/V (12.3 M) solution that contain a stabilizer such as methanol(10-15 %). The 37 % formaldehyde solution is used for denaturing agarose gel containing formaldehyde. Forinstance, Sigma-Aldrich supplies formaldehyde solution, 36.5-38 % in water, for molecular biology, whichcontains 10-15 % methanol.
10 × MOPS buffer *0.2 M
MOPS
20 mM Sodium acetate
10 mM EDTA (pH 8.0)
2. Denaturation of RNA
Prepare denaturated DynaMarker RNA High and RNA to be analysed in a small tube as below.
DynaMarker
RNA High or RNA*2 l
10 × MOPS buffer
2 l
37 % formaldehyde solution
4 lformamide
10 l
200 g/ml ethidum brimode
1 lo
After mixing, heat the RNA solution at 75 C for 3 min, then quickly transfer the tube on ice.
* Required RNA amount depends on experiments. For northern analysis, up to 15 g of RNA is loaded. For
detection of DynaMarker RNA High by ethidum bromide staining, load 0.5-4 g of the marker.
3. Loading and electrophoresis
Add 2 l of 10 × formaldehyde gel-loading buffer* to each RNA solution and return the tube on ice.
Set up the prepared agarose gel containing formaldehyde in a horizontal electrophoresis apparatus submerged in
1 × MOPS buffer. Load the denatured RNA solution to a well. and start electrophoresis. After the tracking dyes
have migrated an appropriate distance through gel, stop the electrophoresis. RNA bands can be seen under UVillumination.
10 × formaldehyde gel-loading buffer*
50 % glycerol
10 mM EDTA (pH 8.0)
0.025 % (w/v) bromophenol blue
0.025 % (w/v) xylene cyanol FF
Reference:
Sambrook, J. and Russell, D.W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, NY.
Related Products:
DynaMarker

DM170

RNA High for Easy Electrophoresis

RNA marker (200-8,000 bases) & RNA Loading Buffer.
RNA sample can be electrophoresed on non-denaturing agarose gel as well as ondenaturing agarose gel with this Loading Buffer.

2

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.6