DescriptionA number of proprietary plasmids are digested to completion with appropriate restriction enzymes to yield 10 bands suitable for use as molecular weight standards for agarose gel electrophoresis. The digested DNA includes fragments ranging from 0.5-10.0 kilobases (kb). The 3.0 kb fragment has increased intensity to serve as a reference band. The approximate mass of DNA in each of the bands is provided (assuming a 0.5 μg load) for approximating the mass of DNA in comparably intense samples of similar size.
Comes supplied with 1 vial of Gel Loading Dye, Purple (6X), no SDS.
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|Gel Loading Dye, Purple (6X), no SDS||25||6X|
Properties and Usage
Effective Size Range500bp to 10,002bp
10 mM Tris-HCl
1 mM EDTA
pH 8.0 @ 25°C
- This ladder was not designed for precise quantification of DNA mass but can be used for approximating the mass of DNA in comparably intense samples of similar size.
- We recommend loading 0.5 μg of 1 kb DNA Ladder diluted in sample buffer.
- All fragments have a 4-base, 5´ overhangs that can be end labeled using T4 Polynucleotide Kinase (#M0201) or filled-in using DNA Polymerase I, Klenow Fragment (#M0210) (1). Use α-[32P] dATP or α-[32P] dTTP for the fill-in reaction.
- 1 kb DNA Ladder is stable for at least 3 months at 4°C.
- For long term storage, store at -20°C. If samples need to be diluted, use TE or other buffer of minimal ionic strength. DNA may denature if diluted in dH20.
- 1X Gel Loading Dye, Purple (6X), no SDS:
3.3mM Tris-HCL (pH 8.0@25°C)
0.02% Dye 1
0.001% Dye 2
- When DNA ladders are run on a polyacrylamide gel, some reference bands may separate due to nucleic acid composition
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual (2nd Ed.). 10.51-10.67.
- Why are there extra bands visible on polyacrylamide gels?
- What are the overhangs on the DNA ladder fragments? Can I end-label them using the T4 polynucleotide kinase (PNK)? What about the Klenow fragment?
- Why are the DNA ladders showing up on my Southern blot? What is the sequence or composition of the ladder bands?
- How can I quantify the amount of DNA in each band of a marker?
- Can I use GelRed with the DNA Ladders from NEB?
- Can I use Midori Green with the DNA Ladders from NEB?
- Can I use SYBR® with the DNA Ladders from NEB?
To make it ready-to-load, dilute in TE buffer instead of water.