Supercoiled Extend DNA Ladder
This product replaces Supercoiled DNA Ladder (NEB #N0472).
Supercoiled Extend DNA Ladder enables accurate sizing of a wide range of supercoiled DNA, including supercoiled plasmids up to 15 kb, and has a convenient reference band at 6 kb.
- Supercoiled DNA molecular weight marker
- Size range: 2 to 15 kb
- Convenient reference band at 6 kb
- Supplied with a vial of Gel Loading Dye, Purple (6X), no SDS (NEB #B7025)
- Suitable for 100 gel lanes
| Catalog # | Concentration | Size |
|---|---|---|
| N0494S | 500 µg/ml | 100 gel lanes (0.1 ml) |
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Product Information
The Supercoiled Extend DNA ladder contains 9 proprietary supercoiled plasmids, ranging in size from 2 to 15 kb, that are suitable for use as supercoiled DNA molecular weight standards in agarose gel electrophoresis. The 6 kb plasmid has an increased intensity to serve as a reference band.
- Recommended gel percentage range: 0.6-1%
- Optimum separation on 0.8%
- Recommended migration conditions: no more than 5V/cm (distance between electrodes), for no more than 60 minutes.
- Comes supplied with 1 vial of Gel Loading Dye, Purple (6X), no SDS
This product is related to the following categories:
- Markers & Ladders
- DNA Markers & Ladders
This product can be used in the following applications:
- DNA Analysis
Reagents Supplied
The following reagents are supplied with this product:
| NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration |
|---|---|---|---|---|---|
| N0494S | Supercoiled Extend DNA Ladder | N0494SVIAL | -20 | 1 x 0.1 ml | 500 µg/ml |
| Gel Loading Dye, Purple (6X), no SDS | B7025SVIAL | 25 | 1 x 1 ml | 6 X |
Properties & Usage
Bases
| Fragment | Mass (ng) | bp |
|---|---|---|
| 1 | 45 | 14996 |
| 2 | 45 | 12000 |
| 3 | 45 | 10002 |
| 4 | 45 | 8001 |
| 5 | 136 | 6000 |
| 6 | 45 | 5000 |
| 7 | 45 | 4000 |
| 8 | 45 | 3001 |
| 9 | 45 | 2017 |
Effective Size Range
2,017bp to 14,996bp
Storage Buffer
10 mM Tris-HCl
1 mM EDTA
pH 8 @ 25°C
Related Products
Companion Products
- Quick-Load® Purple 1 Kb Plus DNA Ladder
- 1 kb Plus DNA Ladder
- Quick-Load® Purple 100 bp DNA Ladder
- Q5® High-Fidelity 2X Master Mix
- Monarch® Spin Plasmid Miniprep Kit
Product Notes
- This product replaces NEB #N0472. Note the change in reference band from 5 kb to 6 kb.
- This ladder may contain some traces of nicked DNA and dimers above the 15 kb plasmid. To minimize nicking of the supercoiled DNA, always use sterile pipette tips and avoid multiple freeze-thaw cycles. The migration of supercoiled plasmids in agarose gels can change depending on agarose concentration, buffer and electrophoresis conditions. Dilute in TE or other buffer of minimal ionic strength. DNA may denature if diluted in dH20.
- The 9 proprietary plasmids are purified, phenol extracted and equilibrated to 10 mM Tris-HCl (pH 8.0) and 1 mM EDTA.
- Centrifuge briefly and mix gently before use. We recommend loading 0.5 µg (1 µl) of the Supercoiled Extend DNA Ladder diluted in sample buffer. 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.
- When sizing supercoiled DNA, avoid running the gel for an extended time at a high voltage, as some supercoiled bands might run as doublets and stretch, which is especially true when using TAE gels and buffer. Running it at a lower voltage and for a shorter time will keep the bands sharper and allow an easier size determination.
- Storage: Dividing in several aliquots is recommended to avoid multiple freeze-thaw cycles.
References
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.). Cold Spring Harbor Laboratory Press.
Protocols, Manuals & Usage
Protocols for this product
- Suggested Loading Protocol for DNA Ladders & Markers
FAQs & Troubleshooting
FAQs
- Can I use Midori Green with the DNA Ladders from NEB?
- Can I use SYBR® and/or GelRed® dyes with the DNA Ladders from NEB?
- Why is my DNA Ladder floating out the wells and Why are there no bands visible in my DNA ladder gel lane?
Quality, Safety & Legal
Quality Control Assays
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 & Change Notifications
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]
- N0494S_v1
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]
- N0494S_v1_10325869
- N0494S_v1_10355834
Safety Data Sheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
- Supercoiled Extend DNA Ladder
- Gel Loading Dye, Purple (6X), no SDS
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.
