DynaMarker, dsRNA Easy Load

Product#: DM185
$320.00
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DynaMarker, dsRNA Easy Load

Cat. No. DM185
Storage -80°C
Size 125 μl, about 25 loadings
Range 10 - 1,000 base of RNA
Loading 5 μl is recommended for loading to a well 
Storage condition This product is shipped on dry ice. Upon receipt, store it at - 80°C. Repeated freeze/thaw cycles should be avoided.


Description

The DynaMarker dsRNA Easy Load is supplied in a ready-to-load mixture of loading dye (containing Tris-HCl buffer, glycerol, EDTA sodium salt, sodium chloride, bromphenol blue) and an ideal size marker for determinating sizes of double-stranded RNAs. The DynaMarker dsRNA Easy Load consists of ten double-stranded RNAs, 10, 20, 30, 50, 100, 200, 300, 400, 500 and 1,000 base pairs. In 5 μl of the DynaMarker dsRNA Easy Load, a 20 bp of dsRNA is approximately 50 ng. The DynaMarker dsRNA Easy Load can be visualized by UV light after ethidium bromide staining

Suitable for determinating size of double-stranded RNAs in non-denaturing polyacrylamide gel electrophoresis.

Ten discrete fragments for easy recognition of double-stranded RNA sizes: 10, 20, 30, 50, 100, 200, 300, 400, 500, 1000 bp of dsRNAs.

Amount of 20 bp dsRNA is adjusted to 25 ng/μl approximately.
Two μl of DynaMarker® dsRNA contains about 50 ng (sufficient to detect the band) of 20 bp dsRNA. It is convenient for siRNA analysis.

Product Information in the product(see PDF)describes a method of dsRNA electrophoresis.

DynaMarker dsRNA consists of ten double-stranded RNAs, 10, 20, 30, 50, 100, 200, 300, 400, 500 and 1,000 base pairs. It's supplied in a ready-to-load mixture of loading dye (containing Tris-HCl buffer, glycerol, EDTA sodium salt, sodium chloride, bromphenol blue)
 

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DynaMarker® dsRNA

Electrophoresis profile of DynaMarker® dsRNA (0.5 μg), left lane,dsRNA digested by dicer, right lane, on 7.5 % of acrylamide,1 X TBE buffer as running buffer


Ready-to-load DynaMarker® dsRNA for non-denaturing polyacrylamide gel electrophoresis.

Ready-to-load dsRNA marker consisted of ten discrete fragments for easy recognition of RNA sizes: 10, 20, 30, 50, 100, 200, 300, 400, 500, 1000 bp.

Amount of 20 bp dsRNA is adjusted to 10 ng/μl approximately.
Five μl of DynaMarker® dsRNA Easy Load contains about 50 ng of 20 bp dsRNA (sufficient to detect the band). It is convenient for siRNA analysis.

Supplied 6 × dsRNA Loading Buffer is for easy sample preparation to run dsRNA samples on a non-denaturing polyacrylamide gel electrophoresis.

Product Information in the product(see PDF)describes a method of dsRNA electrophoresis.
Quality Control
After 18 hr incubation of the DynaMarker dsRNA Easy Load at 37 oC, no visible degradation of the marker is observed in 7.5 % polyacrylamide gel electrophoresis

Supplied product
6 × dsRNA Loading Buffer 6 × dsRNA Loading Buffer is used for preparation of dsRNA samples for non-denaturing polyacrylamide gel electrophoresis. One volume of 6 × dsRNA Loading Buffer is added to 5 volumes of sample. The 6 × dsRNA Loading Buffer is RNase free and contains Tris-HCl buffer (pH7.5), glycerol, EDTA sodium salt, bromphenol blue. Store at - 20 o C or - 80 o C.

Note
Even dsRNA is more resistant to RNase than ssRNA, dsRNA is sensitive to degradation by RNase. To avoid damaging the DynaMarker dsRNA, use 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 dsRNA Easy Load on ice and keep it on ice while using.

Recommended usage
The DynaMarker dsRNA Easy Load is manufactured for non-denaturing polyacrylamide gel electrophoresis. As recommended usage, DynaMarker RNA Easy Load is run on 7.5 % polyacrylamide gel as below.

Procedure

1. Preparation of 7.5 % polyacrylamide gel (20 ml gel)
40 % acrylamide : bis solution (29:1) 3.75 ml
 10 × TBE 2.0 ml
 H2O to 20 ml 
 
2. After mixing reagents described above, add 20 μl of TEMED and 160 μl of 10 % ammonium persulfate. Mix quickly and then pour the gel into the mold of a vertical gel apparatus (20 ml is enough gel solution for two 7 cm × 8 cm, thickness 0.1 cm gels). The gel apparatus should be assembled according to the manufacture’s protocol and ready to run with 1 × TBE buffer.

3. Loading and electrophoresis
Prepare dsRNA sample for electrophoresis as below.
1) Size Marker:
DynaMarker dsRNA Easy Load 5 μl
2) Sample to examine:
 dsRNA sample X μl (*)
 Nuclease-free water 4 – X μl
 6 × dsRNA Loading Buffer 1 μl
 Total 5 μl 

(*) 50-500 ng of dsRNAs.

Mix dsRNA solution with 6 × dsRNA Loading Buffer in a tube as above. Load the mixture onto a well of 7.5 % polyacrylamide 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 μg/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 Harbor Laboratory Press, Cold Spring Harbor, NY.
 
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