Applied Biosystems™ BigDye XTerminator™ Purification Kit

The BigDye XTerminator™ Purification Kit is a rapid and efficient purification system designed to remove unincorporated BigDye™ terminators and salts from DNA sequencing reactions. This method eliminates dye blobs and significantly enhances sequencing accuracy. The process requires only two reagents, making it faster, simpler, and more reliable than traditional ethanol precipitation methods.

✔ Cleanup in under 40 minutes with less than 10 minutes of hands-on time.

✔ Compatible with manual and automated workflows.

✔ Stable for 48 hours at room temperature or 10 days at 4°C.

Catalog Numbers & Quantities

Key Features & Benefits

✔ Complete Dye Blob Removal – Eliminates unincorporated dye terminators, preventing artifacts in sequencing data.

✔ Fast & Simple – Requires only two reagents, reducing workflow complexity.

✔ Improved Long & Short Fragment Recovery – Increases sequence read length and enhances short fragment sequencing quality.

✔ Enhanced Sample Stability – Purified samples are stable for 48 hours at room temperature or 10 days at 4°C.

✔ Automatable Workflow – Compatible with manual or high-throughput automated protocols.

✔ Reliable & Reproducible – Produces high-quality sequencing results with greater consistency.

How It Works: The Simple Purification Process

Traditional Ethanol Precipitation vs. BigDye XTerminator™ Purification

Traditional purification methods involve multiple reagents, centrifugation, and liquid transfers. The BigDye XTerminator™ Purification Kit streamlines the process using just two reagents:

1. XTerminator™ Solution – Captures unincorporated dye terminators and removes free salts.
2. SAM™ Solution – Enhances the purification process and stabilizes the sequencing reaction.

Step-by-Step Protocol

1. Perform Cycle Sequencing – Use BigDye™ Terminators in a standard sequencing reaction.
2. Add Reagents – After sequencing, briefly centrifuge the reaction plate and add SAM™ Solution.
3. Add XTerminator™ Solution – Pipette the correct volume of XTerminator™ Solution into each well.
4. Vortex & Centrifuge – Vortex the plate for 30 minutes to ensure proper mixing, then briefly centrifuge.
5. Run the Reaction Plate – Load the purified sequencing reactions onto an Applied Biosystems™ DNA Analyzer and start the sequencing run.

Performance Advantages

1. Excellent Long Fragment Recovery

• Maintains long-read sequence length without compromising data quality.
• Example: The DNA Technology Unit at NRC-PBI (Canada) achieved Q20 base read = 1044 bp.

2. Improved Short Fragment Recovery

• Outperforms ethanol-based purification, which often results in dye blob interference.
• Example: NRC-PBI demonstrated cleaner short fragment sequencing using BigDye XTerminator™.

3. Increased Sample Stability

• Room temperature stability (48 hours) ensures flexibility in instrument loading.
• Cold storage stability (10 days at 4°C) allows re-runs without reprocessing.

4. Compatible with Multiple Sequencing Platforms

• Optimized for:
  • Applied Biosystems™ 3100, 3100-Avant, 3130/3130xl, 3730/3730xl Genetic Analyzers
  • Works with BigDye™ Terminator v1.1, v3.1, and BigDye™ Direct Cycle Sequencing Kits

Specifications & Technical Details

Contents & Storage

✔ Kit Includes:

• XTerminator™ Solution (store at 4°C)
• SAM™ Solution (store at room temperature or 4°C)

✔ Storage Conditions:

• XTerminator™ Solution: 4°C
• SAM™ Solution: Room temperature (or 4°C)
• Shipping Condition: Wet Ice

Instrument Compatibility & Software Support

✅ Compatible with:

• Applied Biosystems™ 3100, 3100-Avant, 3130, 3130xl, 3730, 3730xl
• BigDye™ Terminator v1.1, v3.1, and BigDye™ Direct Cycle Sequencing Kits

📌 Run Modules Available:

• Software modules for BigDye XTerminator™ are available for Windows 2000 & Windows XP operating systems.
• Applied Biosystems provides downloadable sequencing run modules for supported genetic analyzers.

Ordering Information

Regulatory & Compliance Information

• For Research Use Only (RUO) – Not intended for diagnostic procedures.
• Manufactured under strict quality control standards for consistent and reproducible sequencing results.

The BigDye XTerminator™ Purification Kit is an efficient, fast, and reliable solution for removing unincorporated dye terminators and salts from sequencing reactions. By simplifying the workflow and improving sequence read quality, it is the ideal purification method for Sanger sequencing labs, research facilities, and high-throughput sequencing centers.