Product Name :Code No. :Range :
Size :

DynaMarker

Small RNAⅡ

DM192
20-100 base of RNA
30 loadings, 30 μl ( approximately 6-8 μg)
This product is research use only

Description :The DynaMarker Small RNAⅡ consists of five single-stranded RNAs
(ssRNA). The 20, 30, 40 and 50 bases RNAs are synthesized bychemically (non phosphorylated). The 100 bases RNA is synthesized
by in vitro transcription. The DynaMarker® Small RNAⅡ is suitable fordeterminating size of small-size ssRNA in denaturing polyacrylamide
gel electrophoresis. The DynaMarker Small RNAⅡ can be visualized byethidium bromide staining.

base－ 100

－ 50－ 40
－ 30

Storage buffer :10 mM Tris-HCl (pH 8.0) buffer containing 1 mM EDTA
(Ammonium acetate is slightly contained)－ 20

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

Quality Control :After 18 hrs incubation of the DynaMarker Small RNAⅡ at 37 oC, novisible degradation of the marker is observed in 12.5 % polyacrylamide/ 7.5 M urea gel electrophoresis

Small RNAⅡElectrophoresis profile of
DynaMarker Small RNA Ⅱ (1 μl) on

12.5 % of acrylamide, 7.5 M urea gel with
1 × TBE buffer as running buffer

Note :・The DynaMarker Small RNA II is not prepared for estimating of RNA amount.
・The DynaMarker Small RNA II should be run on 10-20% denaturing polyaceylamide gel for sizing RNAs.
・RNA is very sensitive to degradation by nucleases. To avoid damaging the DynaMarker Small RNAⅡ, use
extreme 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 be high grade and nucleasefree. To use, thaw the DynaMarker Small RNAⅡ on ice and keep it on ice while using.

1

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.6

Recommended usage :・The DynaMarker Small RNAⅡis suitable for 10-20% denaturing polyacrylamide gel electrophoresis. As
recommended usage, DynaMarker Small RNAⅡcan be run on 12.5 % polyacrylsmide / 7.5 M urea gel asbelow.
・Procedure1.
Preparation of 40 % Acrylamide : bis solution
Acrylamide
190 g
N, N-methylenebisacrylamide
10 g
ddH2O
to 500 ml
After mixing, filter the solution through a nitrocellulose filter (0.45 m pore size).
2.

Preparation of 12.5 % polyacrylamide / 7.5 M urea gel (20 ml gel)
40 % acrylamide : bis solution
6.25 mlUrea
9.0 g
10 × TBE
2.0 ml
H2O
to 20 ml

After urea is dissolved completely, add 20 l of TEMED and 160 l of 10 % ammonium persulfate. Mix
quickly and then pour the gel into the mold of a vertical gel apparatus (8.7 cm × 6.8 cm, thickness 1 mm).
The gel apparatus should be assembled according to the manufacture’s protocol and ready to run with 1 ×
TBE buffer.
3. Loading and electrophoresis
Mix 5 l of gel loading buffer* with 1 l** of DynaMarker Small RNAⅡor a few g of RNA sample in a small
tube. Heat at 80℃ for 5 min and immediately transfer the tube on ice. Load the mixture onto a well of
12.5 % polyacrylamide / 7.5 M urea gel and start electrophoresis. After the tracking dyes have migrated anappropriate distance through gel, stop the electrophoresis. To stain with ethidium bromide, disassemble theapparatus and transfer the polyacrylamide gel to a gel tray filled with 1 × TBE buffer containing 1.0 g/ml
ethidium bromide. Stained RNA can be visualized using UV transilluminator.
gel loading buffer*
80 %deionized formamide
0.025% (w/v)
bromophenol blue
0.025% (w/v)
xylene cyanol FF
10 mM
EDTA (pH8.0)** The amount is enough to be visualized by ethidium bromide staining.
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

DM253

Prestain Marker for Small RNA Plus

Pre-stained molecular weight marker for a small size RNA. (range: 20 - 100 bases)

2

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.6