Product Class: Nucleic Acid Marker

2-Log DNA Ladder (0.1-10.0 kb)

Now comes supplied with 1 vial of Gel Loading Dye, Purple (6X), no SDS (B7025)
Catalog #SizeConcentrationGel Lanes
N3200S0.1 ml1,000 μg/ml200
N3200L0.5 ml1,000 μg/ml1000

Description

A number of proprietary plasmids are digested to completion with appropriate restriction enzymes to yield 19 bands suitable for use as molecular weight standards for agarose gel electrophoresis. This digested DNA includes fragments ranging from 100 bp to 10 kb. The 0.5, 1.0 and 3.0 kb bands have increased intensity to serve as reference points. The approximate mass of DNA in each of the bands is provided (assuming a 1.0 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



N3200_thumb

2-Log DNA Ladder visualized by ethidium bromide staining on a 1.0% TBE agarose gel. Mass values are for 1 µg/lane.

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C)Concentration
Gel Loading Dye, Purple (6X), no SDS256X

Properties and Usage

Bases

FragmentMassbp
14010002
2408001
3486001
4405001
5324001
61203001
7402017
8571517
9451200
101221000
1134900
1231800
1327700
1423600
15124500/517
1649400
1737300
1832200
1961100

Effective Size Range

100bp to 10,002bp

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
1 mM EDTA
pH 8.0 @ 25°C

Supporting Documents

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
The following is a list of Material Safety Data Sheets (MSDS) that apply to this product to help you use it safely. The following file naming structure is used to name these document files: [Product Name] MSDS. For international versions please contact us at info@neb.com.
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

Notes

  1. 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.
  2. We recommend loading 0.5-1 μg of the 2-Log DNA Ladder diluted in sample buffer.
  3. All fragments have 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.
  4. 2-Log DNA Ladder is stable for at least 3 months at 4°C.
  5. 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
  6. 1X Gel Loading Dye, Purple (6X), no SDS:
    2.5% Ficoll®-400
    10mM EDTA
    3.3mM
    Tris-HCL (pH 8.0@25°C)
    0.02% Dye 1
    0.001% Dye 2
  7. When DNA ladders are run on an acrylamide gel, some separation of the reference bands may be observed due to variations in the nucleic acid composition of the DNA molecules within that band.

References

  1. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual (2nd Ed.). 10.51-10.67.
  1. Why are there extra bands visible on polyacrylamide gels?
  2. 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?
  3. Why are the DNA ladders showing up on my Southern blot? What is the sequence or composition of the ladder bands?
  4. How can I quantify the amount of DNA in each band of a marker?
  5. Can I use GelRed with the DNA Ladders from NEB?
  6. Can I use Midori Green with the DNA Ladders from NEB?
  7. Can I use SYBR® with the DNA Ladders from NEB?
  1. End-labeling Protocol
  2. Suggested protocol for loading a DNA Ladder/marker

To make it ready-to-load, dilute in TE buffer instead of water.