Product Name :DynaMarker

Code No. :DM185

Range :

10 - 1,000 base of RNA

Size :

125 l,Loading :5 l is recommended for loading to a well

dsRNA Easy Loadabout 25 loadings
This product is research use only

Description :The DynaMarker dsRNA Easy Load is supplied in a ready-to-load
mixture of loading dye (containing Tris-HCl buffer, glycerol, EDTA

origin(well)sodium salt, sodium chloride, bromphenol blue) and an ideal size

bpmarker for determinating sizes of double-stranded RNAs. The

1000500
400300200

DynaMarker

dsRNA Easy Load consists of ten double-stranded RNAs,

10, 20, 30, 50, 100, 200, 300, 400, 500 and 1,000 base pairs. In 5 lof theDynaMarker

dsRNA Easy Load, a 20 bp of dsRNA isapproximately 50 ng. The DynaMarker dsRNA Easy Load can bevisualized by UV light after ethidium bromide staining.

10050Storage condition :30Store at - 80°C.
Repeated freeze/thaw cycles should be avoided.

20Quality Control : After 18 hr incubation of the DynaMarker dsRNA
Easy Load at 37 oC, no visible degradation of the marker is observed

10in 7.5 % polyacrylamide gel electrophoresis
Supplied product :6 × dsRNA Loading Buffer

Electrophoresis profile ofdsRNA Easy Load(5 l) on 7.5 % ofpolyacrylamide,
1 × TBE buffer as runningbuffer

DynaMarker

6 × dsRNA Loading Buffer is used for preparation of dsRNA
samples for non-denaturing polyacrylamide gel electrophoresis. One
volume of 6 × dsRNA Loading Buffer is added to 5 volumes ofsample. The 6 × dsRNA Loading Buffer is RNase free and contains
Tris-HCl buffer (pH7.5), glycerol, EDTA sodium salt, bromphenol
blue. Store at - 20 oC or - 80 oC.
Note :

Even dsRNA is more resistant to RNase than ssRNA, dsRNA is sensitive to degradation by RNase. To avoid
damaging the DynaMarker dsRNA, use care during 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 product should be high grade andnuclease-free. To use, thaw the DynaMarker dsRNA Easy Load on ice and keep it on ice while using.

1

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.3

Recommended usage :The DynaMarker dsRNA Easy Load is manufactured for non-denaturing polyacrylamide gel electrophoresis. As
recommended usage, DynaMarker RNA Easy Load is run on 7.5 % polyacrylamide gel as below.
Procedure
1. Preparation of 7.5 % polyacrylamide gel(20 ml gel)

40 % acrylamide : bis solution (29:1)

3.75 ml

10 × TBE

2.0 ml

H2O

to 20 ml

2. After mixing reagents described above, add 20 l of TEMED and 160 l of10 % ammonium persulfate. Mix quickly and then pour the gel into the moldof a vertical gel apparatus (20 ml is enough gel solution for two 7 cm × 8 cm,thickness 0.1 cm gels). The gel apparatus should be assembled according to
the manufacture’s protocol and ready to run with 1 × TBE buffer.
3. Loading and electrophoresis
Prepare dsRNA sample for electrophoresis as below.
1)

Size Marker:

2)

Electrophoresis of5 l

DynaMarker dsRNA at the left lane anddsRNA sampleX l (*)

Nuclease-free water

4 – X la long dsRNA digested by dicer at
the right lane, on 7.5 % ofpolyacrylamide, 1 × TBE buffer

6 × dsRNA Loading Buffer

1 lDynaMarker

dsRNA Easy LoadSample to examine:

Total 5 l(*) 50-500 ng of dsRNAs.
Mix dsRNA solution with 6 × dsRNA Loading Buffer in a tube as above. Load the mixture onto a well of 7.5 %polyacrylamide gel and start electrophoresis. After the tracking dyes have migrated an appropriate distancethrough gel, stop the electrophoresis. To stain with ethidium bromide, disassemble the apparatus and transfer thepolyacrylamide gel to a gel tray filled with 1 × TBE buffer containing 10 g/ml ethidium bromide. Stained RNA
can be visualized using UV transilluminator.

Reference:Sambrook, J. and Russell, D.W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor
Laboratory Press, Cold Spring Harbor, NY.

2

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.3