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An improved method for determination of gene copy numbers in transgenic mice by serial dilution curves obtained by real-time quantitative PCR assay

Authors
Journal
Journal of Biochemical and Biophysical Methods
0165-022X
Publisher
Elsevier
Publication Date
Volume
64
Issue
2
Identifiers
DOI: 10.1016/j.jbbm.2005.05.006
Keywords
  • Real-Time Quantitative Pcr
  • Taqman
  • Gene Copy Number
  • Zygosity
  • Transgenic Mice
Disciplines
  • Design

Abstract

Abstract Precise characterization of transgene insertion is necessary for phenotype interpretation of transgenic animals. To check for the presence of deletions, estimate the number of inserted transgene copies, and in addition, identify the zygosity of transgenic mice, gene copy numbers were determined by real-time quantitative PCR. Instead of correlating tested samples to a single relative standard curve, serial dilution curves were constructed for every mouse sample. A novel statistical approach was designed in which mice with the same copy number were characterized by the adjusted group mean and standard deviation common to the target sequence. This enabled us to characterize the variability of the obtained results, statistically compare different groups of mice and estimate precision and limits of the applied method.

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