GFP+ bead-bound nuclei were resuspended in buffer RLT for RNA extraction using the RNeasy Micro Kit, along with the recommended on-column DNase digestion (Qiagen 74004). RNA quality and concentrations were measured using an Agilent Bioanalyzer (Agilent RNA 6000 Pico, 5067–1513) and samples having RIN Scores higher than 7 were used. Total RNA was converted to cDNA and amplified using the Ovation RNA-Seq System V2 (Nugen 7102). All samples underwent SPIA Amplification (single primer isothermal amplification) (Ovation RNA-Seq System V2, NuGen, 7102–08) prior to library preparation (NEBNext Ultra DNA Library Prep Kit for Illumina, NEB, 7370S). Three independent samples for each condition were barcoded using the NEBNext Index primers (NEB E7335S), pooled together as one sample, and run on the Illumina platform HiSeq2500 (Rapid Mode-2×50bp) (Macrogen). Next-generation RNA-sequencing generated 40–80 million paired 50 base reads per library.
Hiseq2500 rapid mode 2 50bp
Manufactured by Illumina
The HiSeq2500 (Rapid Mode-2×50bp) is a high-throughput sequencing instrument developed by Illumina. It is capable of generating up to 300 million sequencing reads per run, with read lengths of 2x50 base pairs. The instrument operates in a rapid mode, allowing for faster turnaround times compared to standard sequencing modes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using hiseq2500 rapid mode 2 50bp
1
RNA-Seq Library Preparation and Sequencing
2
RNA-Seq Library Preparation and Sequencing
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GFP+ bead-bound nuclei were resuspended in buffer RLT for RNA extraction using the RNeasy Micro Kit, along with the recommended on-column DNase digestion (Qiagen 74004). RNA quality and concentrations were measured using an Agilent Bioanalyzer (Agilent RNA 6000 Pico, 5067–1513) and samples having RIN Scores higher than 7 were used. Total RNA was converted to cDNA and amplified using the Ovation RNA-Seq System V2 (Nugen 7102). All samples underwent SPIA Amplification (single primer isothermal amplification) (Ovation RNA-Seq System V2, NuGen, 7102–08) prior to library preparation (NEBNext Ultra DNA Library Prep Kit for Illumina, NEB, 7370S). Three independent samples for each condition were barcoded using the NEBNext Index primers (NEB E7335S), pooled together as one sample, and run on the Illumina platform HiSeq2500 (Rapid Mode-2×50bp) (Macrogen). Next-generation RNA-sequencing generated 40–80 million paired 50 base reads per library.
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