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COSMCPREP HIGH AND LOW COPY PLASMID PURIFICATION

CosMCPrep is a versatile SPRI-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 and produces high quality templates. The single protocol, ease of automation and high quality results make CosMCPrep an ideal solution for all template purification needs.

COSMCPREP FEATURES
  • High quality plasmid purification
  • Single protocol for all template types
  • Fully automated protocols
  • No centrifugation or filtration steps
  • Superior downstream sequencing results

    FLEXIBLE PURIFICATION

    CosMCPrep can be used for purification of:
  • BACs
  • Cosmids
  • Fosmids
  • Low copy plasmids
  • High copy plasmids





    LOW COPY PLASMID PURIFICATION

    Purification of low copy plasmids can be challenging due to the low numbers per cell. CosMCPrep enables efficient, fully-automated purification of low copy plasmids such as fosmids (Figure 1) and BACs. CosMCPrep-purified plasmids are high quality and result in improved sequencing performance (Figure 2). Figure 3 shows a BAC end sequence with a Phred20 read length of 700bp.

    HIGH COPY PLASMID PURIFICATION
    CosMCPrep successfully purifies plasmids derived from a number of vectors propagated in a variety of bacteria, including DH5a, DH10B and JM101. Figure 4 shows the products of a CosMCPrep purification from nine plasmid clones. The high quality of CosMCPrep-purified plasmids is demonstrated by the quality of sequencing results, such as pass rates in excess of 90% and Phred20 values routinely above 650bp.
    COSMCPREP AND THE RICKETTSIA GENOME PROJECT
    One of CosMCPreps unique features is the ability to use a single reagent system to purify a variety of template types. CosMCPrep was exclusively used as the template purification method for the Rickettsia Genome Sequencing Project performed at Agencourt. The complete automation of CosMCPrep enabled Agencourt to complete all sequencing in under four days.
    SEQUENCING STRATEGY
    Genomic DNA was cloned into both fosmid (pEPI-FOS57.5kb single copy per cell vector; 35-40kb inserts) and low copy plasmid vectors (pBT3.2kb plasmid; 5-10 copies per cell) for sequencing and functional genomic studies. More than 30,000 sequencing reactions were run from the purified templates. The low copy plasmid sequencing results, shown in Figure 5, had average Phred20 bases in excess of 640 and an average pass rate of 90%.

    COMPLETE COSMCPREP AUTOMATION
    CosMCPreps streamlined protocol does not require the labor-intensive process of cellular debris removal making CosMCPrep a fully-automated plasmid purification system. Automation of CosMCPrep on medium or high throughput robotic platforms has proven to quickly and cost effectively purify high quality plasmids (Figure 6).









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