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SPRINTPREP SINGLE STEP PLASMID PURIFICATION

SprintPrep is an efficient SPRI-based plasmid purification system that can be completed in less than 15 minutes. Unlike most traditional plasmid purification chemistries, SprintPrep isolates plasmid DNA directly from overnight cultures, which can improve the turnaround time of a sequencing pipeline by as much as a full day. SprintPreps simple protocol requires no sample transfers, no centrifugation or filtration and is easily automated in both 96 or 384-well formats.

SPRINTPREP FEATURES

EASE OF USE
  • No cell pelleting, resuspension or
  • lysate clearing
  • Perform the prep in the culture plate

    SPEED
  • 15 minute prep
  • Streamlines sequencing pipeline


  • HIGH QUALITY SEQUENCING RESULTS
  • >650 Phred20 read lenghts
  • >90% pass rates

    FLEXIBILITY
  • 96 or 384-well formats
  • Manual or fully automated protocol



    UNSURPASSED EASE
    SprintPrep generates sequence data in under 30 hours with only a single plate transfer (Figure 1). Traditional filtration-based plasmid purification and rolling circle amplification reactions require multiple plate transfers or labor-intensive cell pelleting and lysate clearing steps.



    SIMPLE PLASMID PURIFICATION
    Unlike other purification technologies that require sample transfer and labor intensive centrifugation or filtration steps, SprintPrep isolates plasmid DNA directly from overnight cultures. SprintPrep was validated using a variety of plasmids including pOT, pDNR, pFLCIII, pZErO and pBluescript. Figure 2 demonstrates the uniform purification of a variety of plasmids.



    REPRODUCIBLE AND ROBUST 384-WELL PURIFICATION

    SprintPrep produces uniform plasmid purification without the need for normalization of cell number (Figure 3). 384 individual plasmids were purified from 150 l of cultures grown for 16 hours. Despite well to well variations in initial cell concentration, all 384 purifications produced consistent plasmid yields.



    HIGH QUALITY SEQUENCING RESULTS
    SprintPreps direct purification protocol produces superior sequencing results. Plasmid DNA purified using SprintPrep routinely produces Phred20 read lengths greater than 650bp. Figure 4 summarizes results for a Chimpanzee sequencing project conducted at Agencourt. The average Phred20 read length for over 11,000 reads was 624bp. The high quality and uniform quantity of purified DNA leads to higher and more even sequencing signals.



    SPRINTPREP AND THE MAMMALIAN GENE COLLECTION (MGC)
    SprintPrep has been validated on more than 15,000 cDNA clones as part of the Mammalian Gene Collection. Purified plasmids were sequenced using 30ng of template DNA in a 1/16th BigDye v3.1 terminator reaction. Unincorporated dye-terminators were removed using CleanSEQ and sequences read on ABI 3700 and 3730xl's. Overall project results are summarized in the table below and a typical 3700 electropherogram is shown in Figure 5. Agencourt has submitted over 600,000 cDNA sequences to GenBank as part of the MGC and continues to do large-scale cD NA library screening as part of this initiative.



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