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* key references chosen by Carl Wittwer

Basics of the Technology
  1. *Wittwer CT, Ririe KM, Andrew RV, David DA, Gundry RA, Balis UJ: The LightCycler: a microvolume multisample fluorimeter with rapid temperature control. Biotechniques. 22, 176-181 (1997).

  2. *Wittwer CT, Herrmann MG, Moss AA, Rasmussen RP: Continuous fluorescence monitoring of rapid cycle DNA amplification. Biotechniques. 22, 130-131, 134-138 (1997).

  3. *Ririe KM, Rasmussen RP, Wittwer CT: Product differentiation by analysis of DNA melting curves during the polymerase chain reaction. Anal Biochem. 245, 154-160 (1997).

  4. *Lay MJ, Wittwer CT: Real-time fluorescence genotyping of factor V Leiden during rapid-cycle PCR. Clin Chem. 43, 2262-2267 (1997).

  5. *Morrison TB, JJ Weis, and CT Wittwer.  Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification.  BioTechniques, 954-962, 1998.

  6. Crockett AO, Wittwer CT: Fluorescein-labeled oligonucleotides for real-time PCR: using the inherent quenching of deoxyguanosine nucleotides. Anal Biochem. 290, 89-97 (2001).

Bernard PS, CT Wittwer.  Homogeneous amplification and variant detection by fluorescent hybridization probes.  Clin. Chem., 46:147-148, 2000.


Advanced Techniques

  1. Bernard PS, GH Pritham, CT Wittwer.  Color multiplexing hybridization probes using the apolipoprotein E locus as a model system for genotyping.  Anal. Biochem., 273:221-228, 1999.

  2. Elenitoba-Johnson, KSJ, SD Bohling, CT Wittwer and TC King.  Multiplex PCR by multicolor fluorimetry and fluorescence melting curve analysis.  Nature Med., 7:249-253, 2001.


  1. Bernard PS, MJ Lay, CT Wittwer.  Integrated amplification and detection of the C677T point mutation in the methylenetetrahydrofolate reductase gene by fluorescence resonance energy transfer and probe melting curves.  Anal. Biochem., 255:101-107, 1998.

  2. Bernard PS, Ajioka RS, Kushner JP, Wittwer CT: Homogeneous multiplex genotyping of hemochromatosis mutations with fluorescent hybridization probes. Am J Pathol. 153, 1055-1061 (1998).

  3. Lyon E, A Millson, T Phan, and CT Wittwer.  Detection and identification of base alterations within the region of factor V Leiden by fluorescent melting curves.  Mol. Diag., 3:203-210, 1998.

  4. Gundry CN, PS Bernard, MG Herrmann, GH Reed, and CT Wittwer.  Rapid F508del and F508C assay using fluorescent hybridization probes, Genetic Testing, 3:365-370, 1999.

  5. Herrmann MG, Dobrowolski SF, Wittwer CT: Rapid beta-globin genotyping by multiplexing probe melting temperature and color. Clin Chem. 46, 425-428 (2000).

  6. Millward, H, W Samowitz, CT Wittwer, PS Bernard.  Homogeneous amplification and mutation scanning of the p53 gene using fluorescent melting curves.  Clin. Chem. 48:1321-1328, 2002.

  7. Seipp MT, Erali M, Wies RL, Wittwer CT.  HLA-B27 typing: evaluation of an allele-specific PCR melting assay and two flow cytometric antigen assays.  Cytometry B Clin Cytom, 2005;63:10-5.

Translocation Detection

  1. Bohling S, TC King, CT Wittwer, and KSJ Elenitoba-Johnson. Rapid simultaneous amplification and detection of the MBR/JH chromosomal translocation by fluorescence melting curve analysis.  Am. J. Pathol., 154: 97-103, 1999.

  2. Bohling S, CT Wittwer, TC King, KSJ Elelitoba-Johnson.  Fluorescence melting curve-based analysis for the detection of the bcl-1/JH translocation in mantle cell lymphoma.  Lab. Invest. 79:337-345, 1999.


  1. Lyon E, A Millson, MC Lowery, R Woods, and CT Wittwer.  Quantification of HER2/neu gene amplification by competitive PCR using fluorescent melting curve analysis, Clin. Chem., 844-851, 2001.

  2. Millson A, Suli A, Hartung L, Kunitake S, Bennett A, Nordberg MC, Hanna W, Wittwer CT, Seth A, Lyon E.  Comparison of Two Quantitative Polymerase Chain Reaction Methods for Detecting HER2/neu Amplification.  J Mol Diagn. 5:184-190, 2003.

Reviews & Book Chapters
  1. *Wittwer CT, K Ririe, R. Rasmussen.  Fluorescence monitoring of rapid cycle PCR for quantification, in Gene Quantification, Ferre, F., ed., Birkhauser, New York, 129-144, 1998.

  2. Pritham GH and CT Wittwer.  Continuous fluorescent monitoring of PCR. J. Clin. Lig. Assay, 21(4):404-412, 1998.

  3. DeSilva D, CT Wittwer.  Monitoring hybridization during PCR, J. Chromatog. B: 741:3-13, 2000.

  4. *Wittwer CT, Herrmann MG, Gundry CN, Elenitoba-Johnson KS: Real-time multiplex PCR assays. Methods. 25, 430-442 (2001).

  5. Wittwer CT.  Introduction to rapid cycle real-time PCR: methods and applications, in Rapid Cycle Real-Time PCR.  S Meuer, CT Wittwer, K Nakaguwara, eds., Springer, Berlin, 1-8, 2001.

  6. Bernard, PS, CT Wittwer.  Real-time PCR technology for cancer diagnostics.  Clin. Chem. 48:1178-1185, 2002

  7. Wittwer CT.  Introduction to real-time PCR and other homogeneous methods for human DNA analysis, in Molecular Testing in Laboratory Medicine: Selections from Clinical Chemistry, 1998-2001, with Annotations and Updates. DE Bruns, YMD Lo, and CT Wittwer, eds., AACC press, Washington, DC, 3-4, 2002.

  8. Wittwer CT, PS Bernard, AO Crockett, SD Bohling and KSJ Elenitoba-Johnson.  Single nucleotide polymorphism (SNP) genotyping by probe melting curve analysis, in Tumor Markers: Physiology, Pathobiology, Technology and Clinical Applications.  EP Diamandis, HA Fritsche, H Lilja, DW Chan and MK Schwartz, ed., AACC press, Washington, DC, 513-515, 2002.

  9. Gundry CN and CT Wittwer.  SYBR Green I analysis of the trinucleotide repeat responsible for Huntington’s disease.  In:  Rapid Cycle Real-Time PCR – Methods and Applications: Genetics and Oncology.  Dietmaier, C. Wittwer, and Sivasubrananian, (eds), Springer, Berlin, 57-63, 2002.

  10. Chang CC, S Perkins and CT Wittwer.  Measurement of MDR1 gene expression by real-time quantitative RT-PCR using the LightCycler instrument.  In:  Rapid Cycle Real-Time PCR – Methods and Applications: Genetics and Oncology.  Dietmaier, C. Wittwer, and Sivasubrananian, (eds), Springer, Berlin, 199-205, 2002.

  11. *Wittwer CT, Kusukawa N: Real-time PCR, In: Diagnostic Molecular Microbiology; Principles and Applications. Persing DH, Tenover FC, Versalovic J, et al. (Eds.), ASM Press, Washington DC, 71-84, 2004.

  12. Pryor, R, CT Wittwer.  Real Time PCR and Melting Curve Analysis. In:  Clinical Applications of PCR (Methods in Molecular Biology), Lo YMD, Chiu RWK, and Chan KCA, eds.  Humana Press, Totowa NJ, Chapter 3, 19-32, 2006.

  1. *Rapid Cycle Real-Time PCR -- Methods and Applications.  S Meuer, C Wittwer, K Nakaguwara, eds., Springer, Berlin, 2001.

  2. *Rapid Cycle Real-Time PCR – Methods and Applications: Microbiology and Food Analysis.  U. Reischl, C Wittwer, and F. Cockerill, eds., Springer, Berlin, 2002.

  3. *Rapid Cycle Real-Time PCR – Methods and Applications: Genetics and Oncology.  Dietmaier, C. Wittwer, and Sivasubrananian, eds., Springer, Berlin, 2002.

  4. *Rapid Cycle Real-Time PCR – Quantification.  C. Wittwer, M. Hahn, and K Kaul eds., Springer, Berlin, 2004.

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