Agencourt CosMCPrep

The Agencourt CosMCPrep system is a Solid Phase Reversible Immobilization (SPRI) paramagenetic bead-based plasmid purification system that uses a single protocol to purify a variety of high and low copy number template types. This flexible system is easily automated without sacrificing product quality. The single protocol, ease of automation and high quality results make Agencourt CosMCPrep an ideal solution for all your plasmid purification needs.

Application: High and low copy plasmid purification

Downstream Application Techniques: DNA Sequencing

Agencourt CosMCPrep Kit Features

High quality plasmid purification
Superior downstream sequencing results
Automated protocols
Single protocol for all template types

Process Overview

1. Resuspension of pelleted cells in RE1 solution and lysis of bacterial cultures with L2 solution. 2. Addition of N3 solution. Flocculent formed in lysate. 3. Transfer of clear lysate from under flocculent to a fresh 96-well round bottom microtiter plate. 4. Addition of isopropanol and PUR4 solution to bind DNA of interest to magnetic beads. 5. Separation of DNA bound to magnetic beads from contaminants. 6. Washing of DNA with Ethanol. 7. Elution of purified plasmids from magnetic particles using RE1 solution. 8. Transfer samples away from magnetic beads to a new plate.

Flexible Purification

The Agencourt CosMCPrep kit can be used for purification of:

BACs
Cosmids
Fosmids
Low copy plasmid
High copy plasmid

Typical Performance

	Low Copy	High Copy
Culture Volume (2xYT)	800µl	600µl
Yield	300ng-1.0µg	500ng-4.0µg
Phred20 Bases	600	650
Pass Rate¹	88%	90%

1 Passing Read: Average Phred value greater than 20 for bases between 100 bp and 300 bp.

Low and High Copy Plasmid Purification using a Single Protocol

One of the Agencourt CosMCPrep system’s unique features is the ability to use a single process to purify both low and high copy plasmids in a fully automated format. Purified plasmids have exceptional sequencing performance with Phred 20 values routinely greater than 600 base pairs

Sequencing Performance

Agencourt CosMCPrep-purified DNA, whether low or high copy in nature, produces high quality sequencing data. Beckman Coulter Genomics has sequenced greater than 1,000,000 Agencourt CosMCPrep-purified, low copy number plasmids, including fosmids, BACS and low copy plasmids.

Figure 2 summarizes Phred 20 read lengths from a Rickettsia genome sequencing project in which greater than 20,000 sequencing reads were performed on low copy plasmid templates. The average sequencing pass rate exceeded 90%, while Phred 20 read lengths often exceeded 800 base pairs.

Agencourt CosMCPrep Automation

The Agencourt CosMCPrep protocol does not require the labor-intensive step of cellular debris removal, making it easier to automate the plasmid purification process. Automation of the Agencourt CosMCPrep protocol on medium or high-throughput robotic platforms has proven to quickly and efficiently purify high quality plasmids.

High Copy Plasmid Purification

Fosmid Purification

Figure 1: Nine pEPI-FOS5-based fosmids and nine pGEM-based plasmids were purified using identical Agencourt CosMCPrep protocols. Products were separated on a 1.2% agarose gel. A. Lane 1: MW ladder, Lane 2-10: P1-P9 plasmids. B. Lane 1: λ ladder, Lane 2-10: F1-F9 fosmids.

Low Copy Plasmid Sequencing

Figure 2: Low copy plasmids were sequenced using 1/16th BigDye* v3.0. Unincorporated dye-terminators were removed using the Agencourt CleanSEQ kit and sequences read on an ABI 3700.

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 CosMCPrep 4,000 rxn Kit	4,000 preps	L2 Buffer N3 Buffer Pur4 Buffer RE1 Buffer	A29174
Agencourt CosMCPrep 384 rxn Kit	384 preps	L2 Buffer N3 Buffer Pur4 Buffer RE1 Buffer	A37064

Related Products			Product #
Agencourt SPRIPlate 96R - Ring Magnet Plate			A29164

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Beckman Coulter, the stylized logo, Agencourt, AMPure, Biomek, CleanSEQ, SPRI, and SPRIPlate are registered trademarks of Beckman Coulter, Inc.