DynaMarker
Product Name:
Prestain Marker for RNA High
Code No. : DM260
Range: 200-8,000 bases
Size: 180 μl (30 loadings)
Storage: store at -80 ℃base

Description:
The DynaMarker Prestain Marker for RNA High is a visible molecularweight marker for ssRNA, consisting of six colored (blue andpurple) nucleic acids. The six colored bands (apparent molecularweights are 200, 500, 1,000, 2,000, 4,000 and 8,000 bases) aresuitable for monitoring denaturing agarose gel electrophoresis andblotting onto membranes. The DynaMarker Prestain Marker for RNAHigh shows the same mobility as that of the DynaMarker RNA High
(code # DM160) on denaturing agarose gel electrophoresis(>90 % accuracy, see table 1 and figure 2). The DynaMarker Prestain
Marker for RNA High is supplied in a ready-to-use mixture
without requiring heating or the addition of a denaturing agent
before use.

8,000－
4,000－
2,000－1,000－
500－
200－

DynaMarker

Storage buffer:
40 mM MOPS (pH 7.0), 10 mM AcONa, 1 mM EDTA・2Na,
10 % Glycerol

Prestain Marker for RNA High

Figure 1: Electrophoresis profile of DynaMarker® Prestain

Quality Control:
After 24 hrs incubation of the DynaMarker Prestain Marker for RNAHigh at 37 ℃, no visible degradation of the marker is observedin 1 % agarose – 2.2 M formaldehyde gel electrophoresis.

Marker for RNA High (6 μl) on 1 %agarose – 2.2 M forumaldehyde gel / 1 ×
MOPS buffer as running buffer.

Recommended loading volumes:Comb
4 ~ 6 mm
>6 mm

Load volume
4 ~ 6 μl
>6 μl

Note:
・・・・For accurate electrophoretic determination of molecular weights, the DynaMarker RNA High (code #DM160) or DynaMarker RNA Easy Measurement N (code # DM170) should be used.
The migration of the DynaMarker Prestain Marker for RNA High has been optimized to use 0.8 －1.5 % of agarose gel concentration (see table 1).
This product is not for acrylamide gel electrophoresis.
Particularly avoid freeze – thaw cycle.

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BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.5

(1)

(2)

base

RNA Highconcentration of agarose

8000 base
4000
2000
1000
500
200

0.8 %
96.5
98.5
104.4
103.4
106.4
102.7

1.0 %
94.8
100.0
102.8
103.2
102.0
100.0

1.5 %
90.4
92.2
101.6
101.4
103.1108.7base

8,000

8,000
4,000

4,000

2,0002,000(1)

(2)

1,000

1,000
500

500200200

1 % agarose

1.5％ agarose

Table 1: Apparent molecular weights as aFigure 2: Electrophoresis profile of DynaMarker RNA

percentage compared to the DynaMarker

High (1) and DynaMarker Prestain Marker

RNA High (DM160).

for RNA High (2) on 1 % and 1.5 %agarose.

Recommended usage:
The DynaMarker Prestain Marker for RNA High is suitable for monitoring denaturing agarose gelelectrophoresis and blotting onto membrane. One example is shown below:・Electrophoresis and blotting of DynaMarker Prestain Marker for RNA High1) Preparation of 0.8 % agarose – 2.2 M formaldehyde gel
Agarose10 × MOPS
deionized formaldehyde
RNase free watertotal

0.8 g
10 ml
18 ml
72 ml
100 mlDissolve the agarose by boiling in a microwave oven. Cool the solution to 55 ℃ and add 10 ml of
10 × MOPS buffer and 18 ml of deionized formaldehyde. In a fumehood, cast an agarose gel withslots formed by a 4~6 mm comb. Remove the comb, place the gel in the gel tank, and add sufficient
1 × MOPS running buffer to cover to a depth of ~1 mm.
2) Loading and electrophoresis.
Thaw the DynaMarker Prestain Marker for RNA High completely before use. Load your denatured RNA
sample and 6 μl of DynaMarker® Prestain Marker for RNA High (use a 4~6 mm comb) on to a well andrun the gel using 1 × MOPS electrophoresis buffer at 4~5 V / cm.

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BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.5

3) Transfer the DynaMarker Prestain Marker for RNA High and RNA from the gel to positively charged nylon
membranes. (see figure 3 and 4)3-1) Place the gel in RNase-free glass dish, and rinse (for formaldehyde removal) with several changesof sufficient deionized water to cover the gel.
3-2) Add ~10 gel volumes of 3 M NaCl / 10 mM NaOH (transfer buffer) to the dish and soak for 30min.
3-3) Cut a piece of nylon membrane slightly larger than the gel. Soak the membrane and two sheets ofblotting paper of appropriate size in 10× SSC for at least 5 min.
3-4) Place the support (e.g. oblong sponge) in a glass or a plastic dish. Fill the dish with enoughtransfer buffer (soak the support about half-submerged in buffer.).
3-5) Place the gel on the support in inverted position so that it is centered on the wet blotting paper.
3-6) Place the wet nylon membrane on top of the gel. (! notice: Remove air bubbles.)3-7) Place the wet blotting paper on the top of the wet nylon membrane. (! notice: Remove airbubbles.)3-8) Cut a stack of paper towels (5~8 cm high), and place the towels on the blotting papers.
3-9) Put a glass plate on the top of the stack and weight it down with an about 400 g weight.
3-10) Allow upward transfer of RNA to occur for 1hr.
3-11) Transfer the membrane to a glass tray containing 6× SSC, and rinse for 5 min.
3-12) Remove the membrane from the 6× SSC and allow excess fluid to drain away. Then dry themembrane on blotting paper for a few minutes.
3-13) Fix the RNA to the membrane with a UV-crosslinker.
3-14) Cut off the marker lane.
3-15) Carry out northern hybridization.

weightglass plate
paper towelnylon membranegelblotting papertransfer buffer
supportFigure 3: Upward Capillary Transfer.

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BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.5

(1)(2)

Transfer

BPB

BPB- agarose gel Figure 4:- membrane -Left : Electrophoresis profile of DynaMarker Prestain Marker for RNA High (1) and RNA
sample (2) on 0.8 % agarose – 2.2 M formaldehyde gel / 1 × MOPS buffer as
running buffer.
Right : Blotting of (1) and (2) onto nylon membrane.

References:・ Joseph Sambrook, and David W. Russell (2001) Molecular Cloning: A Laboratory Manual, 3rd ed.,Cold Spring Harbor Laboratory Press.
・ Frederick M. Ausubel, Roger Brent, Robert E. Kingston, David D. Moore, J. G. Seidman, John A.
Smith, and Kevin Struhl (1994－) Current Protocols in Molecular Biology, John Wiley & Sons, Inc.
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4

BioDynamics Laboratory Inc.
www.biodynamics.co.jp

Ver.1.5