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BigDye XTerminator Purification Kit
Agile, simple purification. It’s in our DNA.
For such a vital step in the sequencing process, you need the highest performance standards and guaranteed results that are not only reproducible, but reliable. Our BigDye XTerminator Purification Kit is a fast, fluid purification method for DNA sequencing reactions that effortlessly removes unincorporated BigDye terminators and salts, paving the way for increased productivity. Drive innovation with this powerful, proven technology, devised by experts, maintained by a global team, and built on a foundation of unrivalled partnership and support.
Although traditional practices such as ethanol precipitation are undeniably lower-cost and readily accessible, efficiency, accuracy, and quality often suffer as a result. These methods can be time-consuming and convoluted, requiring the addition of multiple reagents, along with decanting and centrifuging steps. The BigDye XTerminator Purification Kit, on the other hand, only requires the addition of two reagents, which can be added sequentially or premixed:
- XTerminator Solution—scavenges unincorporated dye terminators and free salts from the post-sequencing reaction
- SAM Solution—enhances the performance of the XTerminator Solution and stabilizes the post-purification reactions
Brochure: Sanger sequencing—Generate high-quality data with our proven workflow
Excellence. You can trust implicitly that sequence length won’t ever be sacrificed, nor will data quality be tainted by dye blobs. Thanks to greater clarity and efficiency, you can progress quickly and confidently to the next stage in your workflow, without any interruptions.
Enjoy peace of mind from start to finish. Sealed reaction plates are stable for 48 hours at room temperature or 10 days at 4°C. Samples can be re-run in case of a power outage or computer malfunction, without having to repeat the experiment. This increases your flexibility when loading a reaction plate on a CE instrument.
While we don’t advise cutting your own pipette tips, because of the inconsistency that may occur as a result of manually cut tips, it can be a short term fix while you wait for your order of our recommended wide-bore pipette tips to arrive. Please note that the tip orifice should be >1.0 mm.
For these, it is only necessary to remove the fluorescently labeled ddNTPs, as they can interfere with the fluorescent sequencing detection. The extension products are single-stranded and their lengths will vary, starting with primer+1, +2, +3, etc. Depending on the method, PCR cleanup methods are intended to remove primers or products below a certain base-pair length, and loss of the smaller sequencing extension products can occur if a PCR purification method is used to purify the sequencing reaction.
Speed and simplicity at your fingertips
Eradicate intrusive salts and terminators quickly and easily with the BigDye Xterminator Purification Kit and prevent their co-injection with dye-labeled extension products. Expect rapid, reliable, and reproducible results, every single time.
- Liquid transfer eliminated with a single plate
- Complete dye blob removal Stabilized samples, primed for analysis
- Cleanup complete in under 40 minutes Less than ten minutes hands-on time End-to-end workflow solution
- Suitable for manual or automated protocols High quality data
- Decades of proven success and published results
Developed by a team of experts and trusted by loyal customers worldwide, the BigDye XTerminator Purification Kit offers superior sequencing data quality and an agile, slick workflow. It swiftly removes unincorporated reaction components, such as salt ions, dye terminator, and dNTPs, in just 40 minutes. A single-plate format negates the need for sample transfer and is compatible with all terminator-sequencing chemistries, reaction volumes and templates, granting you flexibility where it counts. The kit includes:
- BigDye XTerminator Solution—to capture unincorporated dye terminators and free salts from the post cycle-sequencing reaction
- SAM Solution—to improve reagent performance and stabilize the sample after purification
A straightforward protocol that’s built for breakthroughs
Why complicate things? The BigDye Xterminator Purification Kit slots seamlessly into your sequencing workflow with a simple protocol that leaves little margin for error.
- Perform cycle sequencing with BigDye terminators.
- After cycle sequencing, centrifuge the reaction plate briefly, and then pipette the correct volume of SAM Solution into each plate well.
- Pipette the correct volume of XTerminator Solution into each well.
- Vortex the XTerminator Solution bulk container briefly.
- Using a wide-bore pipette tip (recommended wide-bore tips), pipette the correct volume of liquid into each plate well.
- Seal the plate using a heat seal or Clear Adhesive Film (Cat. No. 4306311).
- Vortex for 30 minutes (recommended 96-well microplate vortexers and 384-well microplage vortexers), then centrifuge the reaction plate briefly.
- Place the reaction plate in the Applied Biosystems DNA analyzer. Select run module and run plate.
Self-sufficiency, streamlined
Applied Biosystems supplies downloadable, specialized run modules for use with the BigDye XTerminator Purification Kit and Data Collection Software. These modules are designed for the 3730/3730xl analyzers. The software modules for the BigDye XTerminator Purification Kit are compatible with the Windows 2000 and Windows XP operating systems.
Details at a glance
| BigDye XTerminator Purification Kit |
Test time | 40 minutes |
Format | Kit, which includes:
|
For use with | Sequencing |
Includes | Two bottles |
No. of reactions | 100 |
Stability/storage |
|
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