Product Class: Other

TriDye™ Ultra Low Range DNA Ladder

Product Introduction

  • Convenient, ready-to-load format
  • Size range: 10 bp to 700 bp
  • Suitable for use on both native polyacrylamide and agarose gels.
  • Formulated with TriDye loading dye, containing 3 different tracking dyes
  • Easy to identify reference bands
  • Suitable for 125 to 250 gel lanes

Catalog # Size Concentration
N0558S 1.25 ml

Product Information

Description

TriDye™ Ultra Low Range DNA Ladder is a pre-mixed, ready-to-load molecular weight marker containing three tracking dyes which serve as visual aids to monitor the progress of migration during native agarose and polyacrylamide gel electrophoresis.
 
The DNA ladder contains 13 bands ranging from 10 bp to 700 bp, most of which are generated from the digestion of a proprietary plasmid with restriction enzymes. The 50 bp and 300 bp bands have increased intensity to serve as reference bands. The ladder is suitable for use with for use with both native polyacrylamide and agarose gel electrophoresis.

  • Recommended gel percentage range: 4% TBE agarose, 10% to 20% TBE polyacrylamide

 

Migration of TriDye dyes during electrophoresis*


 Gel  Xylene Cyanol  Bromophenol Blue  Orange G
 4% Agarose  150 bp  20 bp  5 bp
 5% Agarose  100 bp  15 bp  5 bp
 6% PAGE  400 bp  50 bp  15 bp
 10% PAGE  200 bp  30 bp  10 bp
 4-20% PAGE  300 bp  20 bp  10 bp
 20% PAGE  200 bp  20 bp  10 bp

*These migration rates are approximate and variations may occur based on migration conditions (TAE/TBE buffers, buffer pH, voltage etc.).

 



TriDye™ Ultra Low Range DNA Ladder visualized on a 4% TBE agarose gel and on a 20% TBE polyacrylamide gel.
 Mass values are for 0.5 µg/lane.
 Suggested Load: 5 to 10 µl/lane.
This product is related to the following categories:
DNA Markers & Ladders Products
This product can be used in the following applications:
DNA Analysis

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Bases

Fragment Mass (ng) bp
1 49 700
2 46 650
3 28 400
4 63 300
5 28 200
6 11 150
7 14 100
8 21 75
9 70 50
10 10 35
11 16 25
12 19 15
13 125 10

Effective Size Range

10bp to 700bp

Storage Buffer

10 mM Tris-HCl
10 mM EDTA
10% Glycerol
0.006% Xylene Cyanol
0.006% bromophenol blue
0.06% Orange G
pH 8 @ 25°C

Product Notes

  1. Preparation of DNA:
    The double-stranded DNA is digested to completion with appropriate restriction enzymes, phenol extracted and equilibrated in storage buffer.

  2. Loading Recommendations:

    • For polyacrylamide gels, load 5 µl (0.5 µg) of TriDye™ Ultra Low Range DNA Ladder per gel lane.
    • For agarose gels, load 10 µl (1 µg) of TriDye™ Ultra Low Range DNA Ladder per gel lane.
  3. Usage Recommendations: 
    • When using the DNA ladder on agarose gels, we recommend using ethidium bromide at 0.5 ug/ml in both the gel and the migration buffer for proper visualization of the smallest DNA fragments. We also recommend that you do not cast gels exceeding 5mm in thickness, as the smaller DNA fragments might diffuse more easily and may be harder to efficiently stain.
    • We recommend using TBE buffer in the agarose gels and electrophoresis buffers, as TBE buffer allows for a better resolution of the smaller DNA fragments. Fragments below 15 bp might not separate efficiently when using TAE buffer.
    • We do not recommend using GelRed® or SYBR® dyes as precast gel stains, as these dyes might interfere with the migration of the smallest DNA fragments and result in an abnormal band pattern. We recommend using these dyes as post-electrophoresis stains only. Optimization might be required for proper visualization of all DNA fragments. Intensity of some bands might vary.
    • If using wide combs (10 mm) for agarose gel electrophoresis, we recommend loading 15 ul (1.5 ug) of TriDye™ Ultra Low Range DNA Ladder per gel lane for efficient visualization of the smallest DNA fragments.
    • For optimal visualization of all the bands, we recommend exposing the gel to UV without any casting tray whenever possible. Even UV-transparent trays can obscure DNA fragments under UV exposure.
  4. Usage Notes: 
    • When using the DNA ladder on agarose gels, we recommend using ethidium bromide at 0.5 ug/ml in both the gel and the migration buffer for proper visualization of the smallest DNA fragments. We also recommend that you do not cast gels exceeding 5mm in thickness, as the smaller DNA fragments might diffuse more easily and may be harder to efficiently stain.
    • We recommend using TBE buffer in the agarose gels and electrophoresis buffers, as TBE buffer allows for a better resolution of the smaller DNA fragments. Fragments below 15 bp might not separate efficiently when using TAE buffer.
    • We do not recommend using GelRed® or SYBR® dyes as precast gel stains, as these dyes might interfere with the migration of the smallest DNA fragments and result in an abnormal band pattern. We recommend using these dyes as post-electrophoresis stains only. Optimization might be required for proper visualization of all DNA fragments. Intensity of some bands might vary.
    • If using wide combs (10 mm) for agarose gel electrophoresis, we recommend loading 15 ul (1.5 ug) of TriDye™ Ultra Low Range DNA Ladder per gel lane for efficient visualization of the smallest DNA fragments.
    • For optimal visualization of all the bands, we recommend exposing the gel to UV without any casting tray whenever possible. Even UV-transparent trays can obscure DNA fragments under UV exposure.

References

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

Protocols, Manuals & Usage

Protocols

  1. Protocol for loading TriDye™ Ultra Low Range DNA Ladder (NEB #N0558)

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. How much of the TriDye™ Ultra Low Range DNA Ladder should I load?
  2. Why do I only see 12 bands in the Ultra Low DNA Ladder?
  3. Should I use TBE or TAE buffer for my agarose gels?
  4. Can I use the TriDye™ Ultra Low Range DNA Ladder with GelRed® or SYBR® Dyes?
  5. Which polyacrylamide gels can I use?
  6. Why are some of the bands in the DNA Ladder fainter at the bottom of the gel?
  7. Why is the 10bp fragment fainter than the other fragments?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

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]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

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

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.