50 bp DNA Ladder

Description

A number of proprietary plasmids are digested to completion with appropriate restriction enzymes to yield 17 bands suitable for use as molecular weight standards for agarose and polyacrylamide gel electrophoresis. The digested DNA includes fragments ranging from 50-1,350 base pairs. The 200 and 500 base pair bands have increased intensity to serve as reference points.

Comes supplied with 1 vial of Gel Loading Dye, Purple (6X), no SDS.




N3236-new-photo

50 bp DNA Ladder visualized by ethidium bromide staining on a 3% 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
11031350
270916
358766
454700
550650
646600
742550
876500
934450
1031400
1127350
1246300
1357250
14107200
1546150
1669100
178450

Effective Size Range

50bp to 1,350bp

Storage Temperature

-20°C

Storage Conditions

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

Notes

  1. The 50 bp DNA 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. All ends have 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] dCTP or α-[32P] dGTP for the fill-in reaction.
  3. 50 bp DNA Ladder is stable for at least 3 months at 4°C.
  4. 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 be denatured if diluted in dH2O.
  5. We recommend loading 0.5-1.0 µg of 50 bp DNA Ladder diluted in sample buffer.
  6. 1X Gel Loading Dye, Purple, no SDS: 
    2.5% Ficoll®-400
    10 mM EDTA
    3.3 mM Tris-HCl (pH 8.0@25°C)
    0.02% Dye 1
    0.001% Dye 2
  7. Due to the limitations of the acrylamide gel technology, one or two extra bands may be visible on the DNA ladders when run on a polyacrylamide gel.

References

  1. Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, (2nd ed.). 10.51-10.67.

FAQs

  1. Why are there extra bands visible on polyacrylamide gels?
  2. Why is the separation of the lower bands incomplete?
  3. 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?
  4. Why are the DNA ladders showing up on my Southern blot? What is the sequence or composition of the ladder bands?
  5. How can I quantify the amount of DNA in each band of a marker?
  6. Can I use GelRed with the DNA Ladders from NEB?
  7. Can I use Midori Green with the DNA Ladders from NEB?
  8. Can I use SYBR® with the DNA Ladders from NEB?

Tech Tips

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

Protocols

  1. End-labeling Protocol
  2. Suggested protocol for loading a DNA Ladder/marker

Datacards

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.

Selection Charts

Certificate of Analysis

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]

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

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.