Multiplexing of PCR assays in breast cancer analysis /
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| Author / Creator: | Maass, Patrick. |
|---|---|
| Imprint: | Hamburg : Bachelor + Master Pub., 2012. |
| Description: | 1 online resource (54 p.) |
| Language: | English |
| Subject: | |
| Format: | E-Resource Book |
| URL for this record: | http://pi.lib.uchicago.edu/1001/cat/bib/13456820 |
Table of Contents:
- Multiplexing of PCR Assays in Breast Cancer Analysis; Table of Contents; 1 Introduction; 1.1 Breast Cancer; 1.2 RNA Expression Profiling and Prediction; 1.3 Objectives; 2 Materials and Methods; 2.1 Materials; 2.1.1 Instruments and Consumables; 2.1.2 Buffers and Chemicals for Nucleic Acid Purification; 2.1.3 Nucleic Acids, Cell Lines & Tissue Samples; 2.1.4 Synthetic Oligonucleotides; 2.1.5 Chemicals and Kits for PCR; 2.1.6 Software; 2.2 Methods; 2.2.1 General Approach; 2.2.2 Purification of Nucleic Acids; 2.2.3 Creation of a Standard Reference RNA Pool; 2.2.4 Real-Time Kinetic RT-PCR
- 2.2.5 Designing Dually Labeled Primer-Probe Sets2.2.6 Dilution of Primer-Probe Sets; 2.2.7 Setup of Singleplex kPCR Assays; 2.2.8 Evaluation of Different Reporter Dyes; 2.2.9 Controlling of Primer-Probe-Set Performance; 2.2.10 Testing Combinations of Two Primer-Probe Sets (Duplexing); 2.2.11 Testing Combinations of Three Sets (Triplexing); 3 Results; 3.1 Identification of Suitable Reporter Dyes; 3.2 Evaluation of Primer-Probe-Performance; 3.3 Identification of Suitable Duplex Combinations; 3.4 Identification of Suitable Triplex Combinations; 4 Discussion
- 4.1 Statistical Evaluation of Primer Performances4.1.1 Slope; 4.1.2 Efficiency; 4.1.3 Y-Intercept; 4.2 Suitable Reporter Dyes and Quencher; 4.3 Optimization of Assays and kinetic RT-PCR Parameter; 4.4 Conclusion and Outlook; 5 Summary; 6 Literature; 7 Appendix; 7.1 Abbreviations; 7.2 Figures; 7.3 Tables; 7.4 Oligonucleotide Sequences; 7.5 Origins of Breast Cancer Samples for MAVPOOL080623a; 7.6 Acknowledgements