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Accurate and efficient data processing for quantitative real-time PCR using a tripartite plant virus as a model.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Feng, Junli Zeng, Rong Chen, Jishuang |
| Copyright Year | 2008 |
| Abstract | Real-time PCR is becoming a preferred method for quantification of minute amounts of nucleic acids. To achieve the full potential of this technique, accurate and convenient models for post-PCR data analysis are required. In this study, three different models were chosen to quantify the definitive copy numbers of Cucumber mosaic virus (CMV) genomic RNAs using raw fluorescence data of real-time PCR, and equations were proposed to compare their expression levels in virions or in planta. The results, as confirmed by standard curve and Northern blotting methods, show that the expression levels of different genes can be compared more accurately and more efficiently by these equations, especially using theoretical fluorescence (F0) and calibration factors (CF), determined by linear regression PCR (LinRegPCR). Thus, these equations, combined with data analysis by the LinRegPCR method, can greatly enhance the high-throughput quantification ability of real-time PCR, and permit accurate, reliable, and facile investigation of the changes in CMV RNAs accumulation. |
| Starting Page | 807 |
| Ending Page | 818 |
| Page Count | 12 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | http://www.biotechniques.com/multimedia/archive/00003/BTN_A_000112750_O_3689a.pdf?origin=publication_detail |
| PubMed reference number | 18533900v1 |
| Alternate Webpage(s) | https://doi.org/10.2144/000112750 |
| DOI | 10.2144/000112750 |
| Journal | BioTechniques |
| Volume Number | 44 |
| Issue Number | 7 |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Cucumber mosaic virus Fluorescence Linear IgA Bullous Dermatosis Mosaic Organism Mosaic Viruses Northern Blotting Nucleic Acids Plant Viruses Quantitation Reverse Transcriptase Polymerase Chain Reaction Sole of Foot Virion |
| Content Type | Text |
| Resource Type | Article |
