Product Name :Code No. :Range :
Size :

DynaMarker

RNA Low II

DM152
20-500 bases of RNA50 g (72 l), 0.7 mg/ml
This product is research use only

Description :The DynaMarker RNA Low II consists of seven single-stranded RNAs. The
20-base and 50-base RNA are synthesized by chemically (notphosphorylated). The 100, 200, 300, 400 and 500 bases are synthesized
by in vitro transcription. The DynaMarker RNA Low II is suitable fordeterminating size of single-stranded RNAs in denaturing polyacrylamide
gel electrophoresis. The concentration of each RNA (20-500 base) in themarker is approximately 0.1 g/l. It is useful for estimating of RNAamount. The DynaMarker RNA Low II can be visualized by ethidium bromide
staining.

base
500
400
300
200

10050

Storage buffer :10 mM Tris-HCl (pH 8.0) buffer containing 1 mM EDTA

20

Storage condition :Store at -80 oC. Repeated freeze/thaw cycles should be avoided.
Quality Control : After 18 hr incubation of the DynaMarker RNA Low II at
37 oC, no visible degradation of the marker is observed in 5 %polyacrylamide / 8M urea gel electrophoresisDynaMarker

RNA Low II

Electrophoresis profile ofDynaMarker
RNA Low II (0.7 g) on
5 % of acrylamide, 8 M urea gel with
1 × TBE buffer as running bufferNote :RNA is very sensitive to degradation by nucleases. To avoid damaging the DynaMarker RNA Low II, 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 andnuclease-free. To use, thaw the DynaMarker RNA Low II on ice and keep it on ice while using.

Recommended usage :The DynaMarker RNA Low II is suitable for RNA size determining in denaturing polyacrylamide gelelectrophoresis. For one of example, DynaMarker RNA Low II can be 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.

1

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.2

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 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 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 (0.7 g) ** of DynaMarker RNA Low II or a few g of RNA sample
in a small tube. Heat at 80 oC for 3 min and immediately transfer the tube on ice. Load the mixture onto awell of 5 % polyacrylamide / 8M 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 filled with 1 × TBE buffer containing 10
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

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