Friday, November 18, 2016

MiRNA Isolation from Plants Rich in Polysaccharides and Polyphenols

MicroRNAs (miRNAs) are a group of 18–22-nucleotide-long noncoding RNAs, which show wide array of roles in various biological and metabolic processes in both animals and plants. They are formed endogenously and have evolutionary conserved sequences. RNA quality is extremely important for sequencing miRNAs using next-generation sequencing platforms. However, isolation and quantification of miRNAs from plant samples are often technically difficult and recovery of miRNAs from total RNA might be problematic. Degradation of RNA weakens the true miRNAs present in the sample, hence it is crucial to use a protocol that effectively retains good integrity miRNAs. In this chapter, we outline few protocols that can be used to maximize the retrieval of good-quality miRNA and total RNA to be used in miRNA analysis for plants rich in polysaccharides and polyphenols.

Sabu, K.K., Nadiya. F. and N. Anjali. 2017. MiRNA Isolation from Plants Rich in Polysaccharides and Polyphenols. In: MicroRNA Profiling. Vol. 1509. Pp 25-36. Sweta Rani (Ed). Springer. ISBN:978-1-4939-6522-9. PMID: 27826915 DOI: 10.1007/978-1-4939-6524-3_4

Genome Size Estimation Using Flow Cytometry in Elettaria cardamomum Maton

The relative 2C genome size and total number of base pairs of small cardamom (Elettaria cardamomum Maton) was determined using flow cytometer. Seven accessions of cardamom were included in the present study. Zea mays L. CE-777 (2C=5.43 pg) was found to be the most suitable reference standard. Samples for analysis were prepared following the two step procedure described by Otto and involving Propidium iodide staining. The fluorescence intensity of 5000 particles was recorded. The mean amount of 2C nuclear DNA of the cardamom sample was calculated as 2.84 pg. Conversion between DNA content and genome size (1 pg DNA=980 Mbp) indicate that the diploid genome size of cardamom is 2783 Mbp. This is the first report of DNA content and genome size in cardamom. Low variation in genome size has been observed for various germplasm accessions including wild, released varieties and landraces.

Anjali, N., Nadiya, F., Shefeek, S. and K. K. Sabu. 2016. Genome Size Estimation Using Flow Cytometry in Elettaria cardamomum Maton. Indian Forester 142 (9), 878-881

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