Abstract
Quantitative (real-time) polymerase chain reaction (PCR) allows the measurement of relative organellar gene copy numbers as well as transcript abundance of individual mitochondrial or plastidial genes. Requiring only minute amounts of total DNA or RNA, the described method can replace traditional analyses like Southern or Northern hybridization which require large amounts of organellar nucleic acids and usually provide only semiquantitative data. Here we describe prerequisites, reaction conditions, and data analysis principles, which should be applicable for a wide range of plant species and experimental situations where comparative and precise determination of gene copy numbers or transcript abundance is requested. Sequences of amplification primers for qPCR of organellar genes from Arabidopsis are provided.
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Acknowledgement
The author thanks R. Reile for providing unpublished data (Fig. 1).
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Weihe, A. (2014). Quantification of Organellar DNA and RNA Using Real-Time PCR. In: Maliga, P. (eds) Chloroplast Biotechnology. Methods in Molecular Biology, vol 1132. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-995-6_14
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DOI: https://doi.org/10.1007/978-1-62703-995-6_14
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-994-9
Online ISBN: 978-1-62703-995-6
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