† The PCR process is covered by patents owned by Roche Molecular Systems, Inc.,
and F. Hoffman-La Roche, Ltd.
* All trademarks are property of their respective owners.
Beckman Coulter, the stylized logo, Agencourt, Biomek, CleanSEQ, SPRI, and SPRIPlate are registered trademarks of Beckman Coulter, Inc.
Agencourt CleanSEQ
The Agencourt CleanSEQ system is a rapid, high performance dye-terminator removal process based on Solid Phase Reversible Immobilization (SPRI) technology. The paramagnetic bead format requires no centrifugation or filtration and is easily performed manually or fully automated for high throughput dye-terminator removal. The Agencourt CleanSEQ system produces sequences with longer Phred 20 read lengths and higher signal intensities than any other purification technology.
Application: Cycle sequencing cleanup
Downstream Application Techniques: DNA Sequencing
Agencourt CleanSEQ Features
- Long Phred 20 read lengths averaging over 700 bps
- Pass rates of 85% or higher
- Increased average signal strength
- Efficient elimination of sequencing reaction contaminants
Process Overview
1. Add Agencourt CleanSEQ reagent and ethanol to sequencing reaction 2. Bind sequencing products to magnetic beads 3. Separate sequencing products from contaminants with magnetic field 4. Wash with ethanol 5. Elute from magnetic particles 6. Transfer away from magnetic beads
How SPRI Technology Saves Money
SPRI technology can significantly reduce sequencing costs. A typical sequencing lab annual budget before and after using Agencourt CleanSEQ is shown below. The overall operating cost can be substantially reduced by using the Agencourt CleanSEQ system. The pie chart on the left shows 200,000 reactions per year, with a dye-terminator volume of 4 µL/reaction and a reagent cost of $0.60/reaction resulting in a $554,040 annual budget cost. The pie chart on the right shows the same number of reactions with the same labor cost. However, by incorporating the Agencourt CleanSEQ reagent and a Beckman Coulter Biomek FX Automated Workstation, the resulting package reduces the annual operating budget to $412,993, which is a savings of $141,047.
Sequencing Lab Annual Budget Analysis
Assumptions: Labor is assumed constant at 35% of overall reagent cost. This will vary from lab to lab and may decrease due to the automation component of the technology. In this example, dye-terminator cost is $4.50/reaction.
Superior Sequencing Data
The Agencourt CleanSEQ system produces high sequencing pass rates and average Phred 20 read lengths greater than 650 base pairs. Results from direct comparison against EtOH precipitation, gel filtration and silica-based magnetic reagents demonstrate the Agencourt CleanSEQ kits superior performance. The process consistently delivers higher signal-to-noise ratios and overall signal intensities, longer Phred 20 read lengths, and is more reproducible than alternative cleanup methods. This enables rapid, more cost effective sequencing.
Increased Signal Performance
Identical 96-well plates were sequenced using BigDye* v3.1 Terminators, dye terminators removed using either Agencourt CleanSEQ or ethanol precipitation and sequences delineated on an ABI 3730xl.
Consistent Performance
Several 96-well plates were sequenced using BigDye v3.1 Terminators and read on an ABI 3730xl. Dye terminators removed using either Agencourt CleanSEQ or DyeEx* kit following manufacturing instructions.
Scalable and Automation-Compatible
The Agencourt CleanSEQ protocol does not require precipitation, filtration or centrifugation. The Agencourt SPRI technology is easily scaled and automation friendly, allowing both high throughput and format flexibility. Agencourt Bioscience has written and optimized software methods for Beckman Coulter’s Biomek Laboratory Automation Workstations.
Ordering Information
To request a quote or make an inquiry use the on-line contact form.
In the United States and Canada call +1 800 361 7780.
Outside the United States and Canada consult the International Distributors list.
| Product | Size | Kit Components | Product # |
|---|---|---|---|
| Agencourt CleanSEQ Kit (8 ml) | 800/1,600 preps1 | Agencourt CleanSEQ Reagent | A29151 |
| Agencourt CleanSEQ Kit (50 ml) | 5,000/10,000 preps1 | Agencourt CleanSEQ Reagent | A29154 |
| Related Products | Product # |
||
|---|---|---|---|
| Agencourt SPRIPlate 96R - Ring Magnet Plate | A29164 |
||
| Agencourt SPRIPlate 384 Magnet Plate | A29165 |
||
| Agencourt SPRI 384-Well Direct Inject Plate | A29166 |
||
| Agencourt SPRI 96-Well Direct Inject Plate | A29173 |
||
1 96 or 384 well format
