Product Name :Code No. :Range :
Size :
Loading :DynaMarker

RNA Low II Easy Load
DM157
20-500 base of RNA
125 l, about 25 loadings
5 l is recommended for loading to a well(0.1 g of each RNA / 5 l )This product is research use onlybase

Description :The DynaMarker RNA Low II Easy Load is supplied in a ready-to-use
mixture of loading dye (containing formamide, EDTA sodium salt,bromphenol blue) and RNAs. It is prepared for denaturing
polyacrylamide gel electrophoresis but not agarose gel elctrophoresis.
The DynaMarker RNA Low II Easy Load has seven single-stranded RNAs,20, 50, 100, 200, 300, 400 and 500 bases. The 20-base and 50-base
RNA are synthesized by chemically (not phosphorylated), others aresynthesized by in vitro transcription. In 5 l of the DynaMarker RNA Low
II, each RNA amount is approximately 100 ng. It is useful forestimating RNA amount approximately. The DynaMarker RNA Low II Easy
Load can be visualized by UV light after ethidium bromide staining.

5004003002001005020Storage condition :Store at -80 oC.
Repeated freeze/thaw cycles should be avoided.

DynaMarker RNA Low IIEasy LoadQuality Control :After 18 hr incubation of the DynaMarker RNA Low II Easy Load at
37 oC, no visible degradation of the marker is observed in 5 %polyacrylamide / 8M urea gel electrophoresis.

Electrophoresis profile of
DynaMarker RNA Low II Easy Load(5 l) on 5 % of acrylamide, 8 M urea
gel with 1 × TBE buffer as runningbuffer

Supplied product : RNA Loading buffer PA
RNA Loading buffer PA is manufactured for denaturing polyacrylamide gel electrophoresis but not agarose gel
elctrophoresis. The loading buffer has a composition of 80 % formamide, 10 mM EDTA sodium salt (pH8.0),
0.025 % bromphenol blue. Store RNA Loading buffer PA at -80 oC. Repeated freeze/thaw cycles should beavoided. It is 1 × to 2 × solution. Use more than one volume of RNA solution.
Note :RNA is very sensitive to degradation by nucleases. To avoid damaging the DynaMarker RNA Low II Easy 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 product should be high grade and nuclease-free. To use, thaw
the DynaMarker RNA Low II Easy Load on ice and keep it on ice while using. For heat denaturation, transfer aliquotof the DynaMarker RNA Low II Easy 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 and safety
glasses. Put a lid tightly at the time of storage.

1

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.2

Recommended usage :The DynaMarker RNA Low II Easy Load is manufactured for denaturing polyacrylamide gel electrophoresis. As
recommended usage, DynaMarker RNA Low II Easy Load is run on 5 % polyacrylamide / 8M urea gel as below.
Effective range of separation of RNAs is about 50 – 500 base in 5 % polyacrylamide / 8M urea gel.
Procedure
1.
Preparation of 40 % Acrylamide : bis solutionAcrylamide
190 g
N, N-methylenebisacrylamide
10 gH2O
to 500 ml
After mixing, filter the solution through a nitrocellulose filter (0.45 m pore size).
2.

Preparation of 5 % polyacrylamide / 8M urea gel (20 ml gel)
40 % acrylamide : bis solution
2.5 mlUrea
9.6 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 (7 cm × 8 cm, thickness 1.0 mm). The gelapparatus 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 l (0.5 – 2 g)
RNA Loading buffer PA5 l - - - over one volume of RNA sample
Mix in a small tube, total 5-7 lTransfer aliquot (5-10 l) of DynaMarker RNA Low II Easy Load to a small tube. Heat RNA mixed with RNALoading buffer PA and DynaMarker RNA Low II Easy Load at 80 oC for 3 min, and transfer the tube on ice
immediately, then load onto a well of 5 % polyacrylamide / 8M urea gel and start electrophoresis. After thetracking dye has 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.
Reference:
Sambrook, J. and Russell, D.W. (2001) Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring HarborLaboratory 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.1.2