AI Article Synopsis
- DNA identification is important for analyzing botanical samples and requires accurate quantification for effective results.
- Real-time quantitative PCR (qPCR) is preferred because it needs less DNA and accurately quantifies diluted samples, making it more efficient than traditional methods.
- This study developed a qPCR assay targeting the rbcL gene in chloroplast DNA from 13 different plant families, demonstrating that qPCR is a reliable tool for DNA quantification in plant samples, which can improve subsequent analyses.
Article Abstract
Species identification from extracted DNA is sometimes needed for botanical samples. DNA quantification is required for an accurate and effective examination. If a quantitative assay provides unreliable estimates, a higher quantity of DNA than the estimated amount may be used in additional analyses to avoid failure to analyze samples from which extracting DNA is difficult. Compared with conventional methods, real-time quantitative PCR (qPCR) requires a low amount of DNA and enables quantification of dilute DNA solutions accurately. The aim of this study was to develop a qPCR assay for quantification of chloroplast DNA from taxonomically diverse plant species. An absolute quantification method was developed using primers targeting the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene using SYBR Green I-based qPCR. The calibration curve was generated using the PCR amplicon as the template. DNA extracts from representatives of 13 plant families common in Japan. This demonstrates that qPCR analysis is an effective method for quantification of DNA from plant samples. The results of qPCR assist in the decision-making will determine the success or failure of DNA analysis, indicating the possibility of optimization of the procedure for downstream reactions.
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| http://dx.doi.org/10.1007/s12033-016-9918-1 | DOI Listing |
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