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REAL Real-Time!

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Recent advances in real-time PCR analysis have led to significant improvements in the accuracy and simplicity of DNA quantification and gene expression analysis. Here we demonstrate the new Bio-Rad iCycler iQ system capabilities. When PCR is conducted on 96 replicates, we achieve a uniformity with a CV of less than 1%, consistent with that of other well-known systems for real-time PCR analysis. We demonstrate the ability to distinguish a two-fold dilution series of human genomic DNA down to 125 genomic equivalents. We also show a wide dynamic range over which quantification is possible, beginning with plasmids or genomic DNA. The iCycler iQ system is designed to work with many detection strategies; here we show the iCycler iQs' ability to employ a variety of techniques, including SYBR Green I, TaqMan® and Molecular Beacons. Finally, the iCycler iQ's unique ability to analyse data at any point within a cycle or dwell time can be a significant advantage when evaluating certain detection chemistries including molecular beacons.

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Boeckman, F., Tan, L. & Hamby, K. REAL Real-Time!. Breast Cancer Res 2, P8.02 (2000) doi:10.1186/bcr174

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Keywords

  • Dilution Series
  • Gene Expression Analysis
  • Dwell Time
  • Full Text
  • Detection Strategy