Total RNA was extracted from fruits at the immature green and breaker stages with three independent biological replicates of each genotype using a Promega ReliaPrep RNA tissue kit according to the manufacturer’s instructions. The RNA concentration was determined with a spectrophotometer (Nanodrop ND-1000; NanoDrop Technologies, Wilmington, DE, United States), RNA quality was assessed with a BioAnalyzer 2,100 (Agilent Technologies), and RNA libraries were constructed following the recommendations of an Illumina Kit (Directional mRNA-Seq Sample Preparation) and sequenced using the Illumina NovaSeq 6,000 System. Each library was sequenced, generating approximately 20 million 150 bp paired-end reads per sample. The raw sequencing reads were analyzed with FastQC1 and, filtered and cleaned using Trimmomatic (Bolger et al., 2014 (link)) (Parameters: ILLUMINACLIP: TruSeq3-PE.fa:2:30:10LEADING:3 TRAILING:3 SLIDINGWINDOW:4:20 MINLEN:50). At least 95% (19.1–27.9 M) of the reads met the quality criteria and were mapped to the tomato reference genome sequence SL3.0 with the ITAG3.2 annotation using STAR v2.4.2. allowing one mismatch (Dobin et al., 2013 (link)), approximately 84% of the reads were uniquely mapped (Supplementary Table 1 ) and were used for statistical analysis.
Reliaprep rna tissue kit
Manufactured by Promega
The ReliaPrep RNA tissue kit is a laboratory equipment product designed for the extraction and purification of RNA from a variety of tissue samples. It is a standardized solution for obtaining high-quality RNA suitable for downstream applications such as reverse transcription and real-time PCR analysis.
Automatically generated - may contain errors
2 protocols using reliaprep rna tissue kit
1
Transcriptome Analysis of Tomato Fruit Ripening
2
Transcriptome analysis of tomato fruit development
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Total RNA was extracted from immature green and breaker stage fruits with three independent biological replicates of each genotype using a Promega ReliaPrep RNA tissue kit according to the manufacturer's instructions. The RNA concentration was determined with a spectrophotometer (Nanodrop ND-1000; NanoDrop Technologies, Wilmington, DE, U.S.A.), RNA quality was assessed with a BioAnalyzer 2100 (Agilent Technologies), and RNA libraries were constructed following the recommendations of an Illumina Kit (Directional mRNA-Seq Sample Preparation) and sequenced using the Illumina NovaSeq 6000 System. Each library was sequenced, generating approximately 20 million 150 bp paired end reads per sample. The raw sequencing reads that were generated were analysed with FastQC (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) and were ltered and cleaned using Trimmomatic 55 (Parameters: ILLUMINACLIP: TruSeq3-PE.fa:2:30:10LEADING:3 TRAILING:3 SLIDINGWINDOW:4:20 MINLEN:50). At least 95% (19.1-27.9 M ) of the reads met the quality criteria and were mapped to the tomato reference genome sequence SL3.0 with the ITAG3.2 annotation using STAR v2.4.2. allowing one mismatch 56 , approximately 84% of the reads were uniquely mapped (Supplementary Table 1) and were used for statistical analysis.
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