title: A simple, accurate and universal method for quantification of PCR.
abstract: BACKGROUND
  Research into gene expression enables scientists to decipher the complex regulatory networks that control fundamental biological processes. Quantitative real-time PCR (qPCR) is a powerful and ubiquitous method for interrogation of gene expression. Accurate quantification is essential for correct interpretation of qPCR data. However, conventional relative and absolute quantification methodologies often give erroneous results or are laborious to perform. To overcome these failings, we developed an accurate, simple to use, universal calibrator, AccuCal.
  
  
  RESULTS
  Herein, we show that AccuCal quantification can be used with either dye- or probe-based detection methods and is accurate over a dynamic range of ≥10(5) copies, for amplicons up to 500 base pairs (bp). By providing absolute quantification of all genes of interest, AccuCal exposes, and circumvents, the well-known biases of qPCR, thus allowing objective experimental conclusions to be drawn.
  
  
  CONCLUSIONS
  We propose that AccuCal supersedes the traditional quantification methods of PCR.
authors.affiliation: Accugen Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. nicolab@accugen.com.au.
authors.forename: Nicky
authors.initials: N
authors.lastname: Boulter
authors.affiliation: ANZAC Research Institute, University of Sydney, Sydney, NSW, 2139, Australia. francia.suarez@sydney.edu.au.
authors.forename: Francia Garces
authors.initials: FG
authors.lastname: Suarez
authors.affiliation: Westmead Millennium Institute, 176 Hawkesbury Road, Westmead, NSW, 2145, Australia. stephen.schibeci@sydney.edu.au.
authors.forename: Stephen
authors.initials: S
authors.lastname: Schibeci
authors.affiliation: Bitfuturistic Solutions, 9623 Lawndale Avenue SW, Lakewood, WA, 98498, USA. tsunderland@bitfuturistic.com.
authors.forename: Trevor
authors.initials: T
authors.lastname: Sunderland
authors.affiliation: Accugen Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. otolhurst@vectusbiosystems.com.au.|Vectus Biosystems Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. otolhurst@vectusbiosystems.com.au.
authors.forename: Ornella
authors.initials: O
authors.lastname: Tolhurst
authors.affiliation: Accugen Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. tegan.hunter@newcastle.edu.au.|Vectus Biosystems Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. tegan.hunter@newcastle.edu.au.
authors.forename: Tegan
authors.initials: T
authors.lastname: Hunter
authors.affiliation: Vectus Biosystems Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. ghodge@vectusbiosystems.com.au.
authors.forename: George
authors.initials: G
authors.lastname: Hodge
authors.affiliation: ANZAC Research Institute, University of Sydney, Sydney, NSW, 2139, Australia. djh@anzac.edu.au.
authors.forename: David
authors.initials: D
authors.lastname: Handelsman
authors.affiliation: ANZAC Research Institute, University of Sydney, Sydney, NSW, 2139, Australia. usimanainen@anzac.edu.au.
authors.forename: Ulla
authors.initials: U
authors.lastname: Simanainen
authors.affiliation: Accugen Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. ehendriks@vectusbiosystems.com.au.|Vectus Biosystems Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. ehendriks@vectusbiosystems.com.au.
authors.forename: Edward
authors.initials: E
authors.lastname: Hendriks
authors.affiliation: Accugen Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. kduggan@vectusbiosystems.com.au.|Vectus Biosystems Pty Ltd, 11 Julius Avenue, North Ryde, NSW, 2113, Australia. kduggan@vectusbiosystems.com.au.
authors.forename: Karen
authors.initials: K
authors.lastname: Duggan
mesh_terms: D000818:Animals; D002138:Calibration; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007963:Leukocytes, Mononuclear; D051379:Mice; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*
chemical_list: D004247:DNA