Total RNA from leaf +1, apical shoot, and fifth and ninth internodes from 6-month-old transgenic and control plants was extracted following the Spectrum Plant total RNA kit protocol (Sigma-Aldrich). Extracts were treated with RQ1 RNase-Free DNase (Promega, USA) according to the manufacturer’s protocol. For each tissue, pooled RNA samples from four biological replicates were used for analysis. One microgram of each RNA sample was used to produce cDNA libraries, which were sequenced using an Illumina HiSeq 2500 by Fasteris Life Science Co. (Geneva, Switzerland). The reads were mapped on Sorghum bicolor reference genome available at Illumina iGenomes (http://support.illumina.com/sequencing/sequencing_software/igenome.html ). The list of differentially expressed genes (DEGs) was identified using an FDR q-value cutoff of 1 × 10−5. The sequence data were deposited in the NCBI SRA database with the accession number SRP132098 (http://www.ncbi . nlm.nih.gov/sra/SRP132098).
Spectrum plant total rna kit protocol
Manufactured by Merck Group
Sourced in United States
The Spectrum Plant total RNA kit protocol is a laboratory equipment product designed to extract total RNA from plant samples. It provides a standardized and efficient method for isolating high-quality RNA from a variety of plant tissues, enabling downstream applications such as gene expression analysis, reverse transcription, and other molecular biology techniques.
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2 protocols using spectrum plant total rna kit protocol
1
Transcriptome analysis of transgenic Sorghum
2
Total RNA Extraction and Purification
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Total RNA was extracted according to the method previously optimised by (Bernáldez, Rodríguez, Rodríguez, Sánchez-Montero & Córdoba, 2017) with the exception of RNA purification was performed using the Spectrum Plant Total RNA Kit protocol (Sigma-Aldrich, USA). RNA was diluted in 50 μL of the elution solution provided by the RNA extraction kit and kept at -80ºC until use for reverse transcription (RT). The RNA concentration and purity (A260/A280 ratio) were determined spectrophotometrically using a 1.5 μL aliquot on the Nanodrop 200 UV-VIS Spectrophotometer (Thermo Scientific, USA).
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