Product Name :DynaMarker

Code No. :DM180

Range :

10 - 1,000 bp of dsRNA

Size :

25 g (100 l), 0.25 mg/ml

dsRNA

This product is research use onlyDescription :The DynaMarker dsRNA consists of ten double-stranded RNAs, 10, 20,30, 50, 100, 200, 300, 400, 500 and 1,000 base pairs. The DynaMarker
dsRNA is an ideal size marker for determinating sizes oforigin(well)

double-stranded RNAs. A twenty bp of RNA band including in theDynaMarker

dsRNA is adjusted to approximately 25 ng/l inconcentration.

TheDynaMarker

dsRNA

ismanufactured

non-denaturing polyacrylamide gel electrophoresis. Thefor

DynaMarker

dsRNA can be visualized by UV light after ethidium bromide

1000
500
400
300
200100staining.
50

Storage buffer :10 mM Tris-HCl (pH 7.5), 50 mM NaCl , 1 mM EDTA sodium salt

3020Storage condition :10Store at - 80°C.
Repeated freeze/thaw cycles should be avoided.

bp
Quality Control : After 18 hr incubation of the DynaMarker dsRNA at
37 oC, no visible degradation of the marker is observed in 7.5 %Electrophoresis profile ofDynaMarker dsRNA (0.5 g),on 7.5 % of polyacrylamide,
1 × TBE buffer as running buffer

polyacrylamide gel electrophoresis

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 marker should behigh grade and nuclease-free. To use, thaw the DynaMarker dsRNA on ice and keep it on ice while using.

1

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.4

Recommended usage :The DynaMarker dsRNA is suitable for RNA size determining in non-denaturing polyacrylamide gelelectrophoresis. For one of example, DynaMarker dsRNA can be run on 7.5 % polyacrylamide gel as below.
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 and160 l of 10 % ammonium persulfate. Mix quickly and then pour thegel into the mold of a vertical gel apparatus (20 ml is enough gel
solution for two 7 cm × 8 cm, thickness 0.1 cm gels). The gelapparatus 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:
Electrophoresis of
DynaMarker
dsRNA
2 l (0.5 g)
DynaMarker dsRNA at the left lane andnuclease-free water
3 la long dsRNA digested by dicer at
6 × gel loading solution
1 lthe right lane, on 7.5 % ofpolyacrylamide, 1 × TBE buffer2) Sample to examine:dsRNA sample
5 l (50-500 ng)
6 × gel loading solution
1 l
Mix dsRNA solution with gel loading solution 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
distance through gel, stop the electrophoresis. To stain with ethidium bromide, disassemble the apparatus
and transfer the polyacrylamide gel to a gel tray filled with 1 × TBE buffer containing 10 g/ml ethidium
bromide. Stained RNA can be visualized using UV transilluminator.
4. Gel loading solution
Gel loading solution is used for non-denaturing polyacrylamide electrophoresis as below.
6 × gel loading solution , example 1
6 × gel loading solution , example 2
0.25 % (w/v) bromophenol blue
0.25 % (w/v) bromophenol blue
0.25 % (w/v) xylene cyanol FF
40 % (w/v) sucrose in H2O
30 % (v/v) glycerol in H2O

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.1.4