title: ExCyto PCR amplification.
abstract: BACKGROUND
  ExCyto PCR cells provide a novel and cost effective means to amplify DNA transformed into competent bacterial cells. ExCyto PCR uses host E. coli with a chromosomally integrated gene encoding a thermostable DNA polymerase to accomplish robust, hot-start PCR amplification of cloned sequences without addition of exogenous enzyme.
  
  
  RESULTS
  Because the thermostable DNA polymerase is stably integrated into the bacterial chromosome, ExCyto cells can be transformed with a single plasmid or complex library, and then the expressed thermostable DNA polymerase can be used for PCR amplification. We demonstrate that ExCyto cells can be used to amplify DNA from different templates, plasmids with different copy numbers, and master mixes left on ice for up to two hours. Further, PCR amplification with ExCyto cells is comparable to amplification using commercial DNA polymerases. The ability to transform a bacterial strain and use the endogenously expressed protein for PCR has not previously been demonstrated.
  
  
  CONCLUSIONS
  ExCyto PCR reduces pipetting and greatly increases throughput for screening EST, genomic, BAC, cDNA, or SNP libraries. This technique is also more economical than traditional PCR and thus broadly useful to scientists who utilize analysis of cloned DNAs in their research.
authors.affiliation: Lucigen Corporation, Middleton, Wisconsin, United States of America.
authors.forename: Vinay
authors.initials: V
authors.lastname: Dhodda
authors.forename: Ronald
authors.initials: R
authors.lastname: Godiska
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Vanwye
authors.forename: David
authors.initials: D
authors.lastname: Mead
authors.forename: Rebecca
authors.initials: R
authors.lastname: Hochstein
authors.forename: Lynne
authors.initials: L
authors.lastname: Sheets
authors.forename: Sarah
authors.initials: S
authors.lastname: Vande Zande
authors.forename: Chris
authors.initials: C
authors.lastname: Niebauer
authors.forename: Douglas L
authors.initials: DL
authors.lastname: Crawford
authors.forename: Marjorie F
authors.initials: MF
authors.lastname: Oleksiak
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014169:Transformation, Bacterial
chemical_list: D004269:DNA, Bacterial