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Flexprep: Scale-flexible rapid plasmid preparation for analysis of recombinant clones

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Abstract

A scale-flexible and cost-efective protocol for plasmid preparation is described to cover miniprep and midiprep scale work in a microcentriguge format for analysis of recombinant clones. this protocol relies on a modified alkaline lysis of Escherichia coli cells and subsequent purification of plasmid DNA with no organic extraction and alcohol precipitation. It can process up to 20 mL of E. coli cells carrying 3–10 kbp plasmid vectors in <10 min. Flexprep delivers sufficient yield and purity of plasmid DNA for routine applications including restriction enzyme digestion and fluorescent automated sequencing.

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References

  1. Birnboim, H.C. and Doly, J. (1979) A rapid alkaline extraction procedure for sequencing recombinant plasmid DNA. Nucleic Acids Res. 7, 1513–1523.

    Article  PubMed  CAS  Google Scholar 

  2. Qiagen (2000) QIAGEN Plasmid Purification Handbook.

  3. Promega (1999) Wizard Plus Midipreps DNA Purification System. Technical Bulletin #173.

  4. Strategene (2000) StrataPrep EF Plasmid Midiprep. Instruction Manual Revision #120004.

  5. Voo, K.S. and Jacobsen, B.M. (1998) Rapid resuspension of pelleted bacterial cells for miniprep plasmid DNA isolation. BioTechniques 24, 240–243.

    PubMed  CAS  Google Scholar 

  6. Ahmed, A. (1994) Double-origin vectors for isolating bidirectional deletions useful in DNA sequence analysis. Gene 141, 71–73.

    Article  PubMed  CAS  Google Scholar 

  7. Boyle, J.S., and Lew, A.M. (1995) An inexpensive alternative to glassmilk for DNA purification. Trends Genet. 11, 8.

    Article  PubMed  CAS  Google Scholar 

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Correspondence to Futoshi Aranishi.

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Aranishi, F. Flexprep: Scale-flexible rapid plasmid preparation for analysis of recombinant clones. Mol Biotechnol 21, 39–41 (2002). https://doi.org/10.1385/MB:21:1:039

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  • DOI: https://doi.org/10.1385/MB:21:1:039

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