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
| Gel | Xylene Cyanol | Bromophenol Blue | Orange G |
| 4% Agarose | 150 bp | 20bp | 5bp |
| 5% Agarose | 100bp | 15bp | 5bp |
| 6% PAGE | 400bp | 50bp | 15bp |
| 10% PAGE | 200bp | 30bp | 10bp |
| 4-20% PAGE | 300bp | 20bp | 10bp |
| 20% PAGE | 200bp | 20bp | 10bp |
*These migration rates are approximate and variations may occur based on migration conditions (TAE/TBE buffers, buffer pH, voltage etc.).
Reagents Supplied
The following reagents are supplied with this product:
| NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
| N0558S | 4 |
| TriDye™ Ultra Low Range DNA Ladder | N0558SVIAL | 4 | 1 x 1.25 ml | 100 µg/ml |
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.
Companion Products
- Quick-Load Purple Low Molecular Weight DNA Ladder
- Low Molecular Weight DNA Ladder
- Monarch® Plasmid Miniprep Kit
- Monarch® DNA Gel Extraction Kit
Notes
- Preparation of DNA:
The double-stranded DNA is digested to completion with appropriate restriction enzymes, phenol extracted and equilibrated in storage buffer. 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.
- 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.
- 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
- 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
- Protocol for loading TriDye™ Ultra Low Range DNA Ladder (NEB #N0558)
Selection Charts
FAQs
- How much of the TriDye™ Ultra Low Range DNA Ladder should I load?
- Why do I only see 12 bands in the Ultra Low DNA Ladder?
- Should I use TBE or TAE buffer for my agarose gels?
- Can I use the TriDye™ Ultra Low Range DNA Ladder with GelRed®or SYBR®Dyes?
- Which polyacrylamide gels can I use?
- Why are some of the bands in the DNA Ladder fainter at the bottom of the gel?
- Why is the 10bp fragment fainter than the other fragments?
Citations
Quality Control Assay
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 Data Sheets
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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.
