This product is an electrophoresis standard consisting of 15 bands between 0.2 and 10 kilobase pairs. Bands are regularly spaced so that sizes are easy to identify. Markers are supplied ready-to-use containing loading dye so that electrophoresis can be monitored.
The Biotinylated DNA Molecular Weight Marker (MB-1302) is used when nucleic acids are to be transferred to a membrane and later detected using an avidin or streptavidin-based system.
| Unit Size | 25 µg |
|---|---|
| Applications | Blotting Applications |
| Solution | 20 mM Tris-HCl, pH 7.5, 2 mM EDTA, 2% Ficoll, 0.08% sodium azide, 0.02% bromophenol blue. |
| Recommended Usage | The banding pattern and band sizes are shown on the left. 5 µl (500 ng) of the ready-to-use ladder were loaded on a 0.5% agarose gel.The gel was run in 1x TBE at 10 V/cm, stained with ethidium bromide (A) and blotted onto nylon membrane using standard methods.B-DNA bands were detected using the UltraSNAP™ Kit (an alkaline phosphatase-streptavidin conjugate followed by chemiluminescent substrate) (B). 1.The marker is provided at a concentration of 100 ng/µl in 20 mM Tris, pH 7.5, 2 mM EDTA, 2%Ficoll, 0.08% sodium azide, 0.02% bromophenol blue.Store the b-DNA marker at 4° C.For long term storage, -20° C is recommended. 2.The recommended loading volume per lane is 5 µl (500 ng). 3.The bromophenol blue tracking dye migrates between the 200 bp and 400 bp bands on a 0.5% agarose gel. 4.The fragments can be visualized in the gel by staining the gel with ethidium bromide.Soak the gel in a solution of 0.5 µg/ml ethidium bromide in distilled water for 30 minutes after electrophoresis and visualize the gel under UV light.Destaining with distilled water for 30 minutes may improvethe visualization of bands.(Caution:Ethidium bromide is a powerful mutagen.Use gloves and other appropriate safety measures with this compound.) 5.Sample DNA, along with the biotinylated marker, can be transferred from the gel to nylon ornitrocellulose membrane and hybridized with biotinylated probe by standard northern or Southern blotting procedures.After blocking (eg. 10x Casein Solution, SP-5020 or Animal-Free Blocker™, SP-5030), the membrane is incubated in the appropriate enzyme-conjugated streptavidin (eg. alkaline phosphatase-streptavidin, SA-5100).Visualization of signal is achieved by incubation in colorimetric substrate (eg. BCIP/NBT, SK-5400) or chemiluminescent substrate(eg. DuoLuX™, SK-6605). |
| Recommended Storage | 2-8 °C, for long term storage -20 °C is recommended |
| Concentration | 100 ng/µl |
| Conjugate | Biotinylated |
10x Casein Solution
Catalog #:
SP-5020
Animal-Free Blocker®, 5X Concentrate
Catalog #:
SP-5030
Features:
Markers are supplied ready-to-use containing loading dye so that electrophoresis can be monitored.
Ready-to-use marker (5 μl or 500 ng) was loaded on a 0.5% agarose gel. The gel was run in 1x TBE at 10 V/ cm, stained with ethidium bromide (left) or blotted onto nylon membrane using standard methods.
