Product Name :Code No. :Range :

DynaMarker

Size :

25 lodings, 125 μl

Loading :5 μl is recommended for loading to a well

Small RNAⅡEasy Load
DM197
20-100 bases of RNA
This product is research use only

Description :The DynaMarker Small RNAⅡEasy Load has five single-stranded RNAs, 20,30, 40, 50 and 100 bases, which is useful for a research of small RNAs.
The 20, 30, 40 and 50 bases RNA are synthesized by chemically (nonphosphorylated). The 100 bases RNA is synthesized by in vitrotranscription. The DynaMarker Small RNAⅡEasy Load is supplied in a readyto-use mixture of loading dye and RNAs (containing formamide, EDTA
sodium salt, bromphenol blue). It is manufactured for denaturing
polyacrylamide gel electrophoresis. The DynaMarker® Small RNA Ⅱ Easy
Load can be visualized by UV light exposure after ethidium bromide
stainingbase

－ 100

－ 50
－ 40
－ 30
－ 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 RNAⅡEasy Load at 37 oC,
no visible degradation of the marker is observed in 12.5 % polyacrylamide/ 7.5 M urea gel electrophoresis.

Small RNAⅡ
Easy LoadElectrophoresis profile ofDynaMarker Small RNAⅡEasy Load (5 μl)on 12.5 % of acrylamide, 7.5 M urea gel
with
1 × TBE buffer as running buffer

Supplied product : RNA Loading buffer PA
RNA Loading buffer PA is manufactured for denaturing polyacrylamide gel electrophoresis. The loading
buffer has a composition of 80 % formamide, 10 mM EDTA sodium salt (pH 8.0), 0.025 % bromphenol
blue. Store RNA loading buffer PA at -80℃. Repeated freeze/thaw cycles should be avoided. It is 1 × to 2× solution. Use RNA Loading buffer PA more than one volume of RNA solution.
Note :・The DynaMarker Small RNAⅡEasy Load is not prepared for estimating of RNA amount.
・The Small RNAⅡEasy Load 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 RNAⅡEasy
Load, 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 nuclease-free. To use, thaw the DynaMarker Small RNAⅡEasy Load on ice and keep it on ice whileusing. For heat denaturation, transfer aliquot of the DynaMarker Small RNAⅡEasy Load to another tube,
then heat it. Avoid repeated heat denaturizing.
‡ Formamide is suspected to be harmful. It is irritate to the eyes and skin. Wear appropriate gloves andsafety glasses. Put a lid tightly at the time of storage
1

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.5

Recommended usage :・ The DynaMarker Small RNA Ⅱ Easy Load is suitable for 10-20% denaturing polyacrylamide gelelectrophoresis. As recommended usage, DynaMarker Small RNAⅡEasy Load can be run on 12.5 %polyacrylsmide / 7.5 M urea gel as below.
・Procedure1.
Preparation of 40 % Acrylamide : bis solution
Acrylamide : bis
190 gN, 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 RNA to be analyzed (for example, RNA transcript) and RNA Loading buffer PA as below.
RNA sample
dried precipitate or 2 μl (0.5-2 μl)
RNA Loading buffer PA
5 μl --- over one volume of RNA sample
Mix in a small tube, total 5-7 μl
Transfer aliquot (5-10 μl) of DynaMarker Small RNAⅡEasy Load to a small tube. Heat RNA mixed withloading buffer PA, and DynaMarker® Small RNAⅡEasy Load at 80℃ for 5 min, and immediately transfer thetube 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 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 filledwith 1 × TBE buffer containing 1.0 μg/ml ethidium bromide. Stained RNA can be visualized using UVtransilluminator.
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 productsDynaMarker

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.1.5