Illumina TruSeq Stranded Total RNA Sequencing

RNA concentration and purity were assessed by absorbance at 260 and 280 nm using NanoDrop One (Thermo Fisher Scientific). RNA was checked for quality using the Agilent RNA 600 nano kit and Bioanalyzer instrument (Agilent Technologies). Aliquots of RNA (1,000 ng) were analyzed using the Illumina TruSeq Stranded Total RNA with Ribo-Zero Gold protocol (Illumina) as described (76 (link)). Samples were cleaned and validated for DNA concentration using the Qubit dsDNA HS assay kit (Invitrogen) and for base pair size and purity using the Agilent High Sensitivity DNA chip and Bioanalyzer instrument. The libraries were subjected to 38-bp paired-end sequencing on a NextSeq500 (Illumina).

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Pendergrast L.A., Lundell L.S., Ehrlich A.M., Ashcroft S.P., Schönke M., Basse A.L., Krook A., Treebak J.T., Dollet L, & Zierath J.R. (2023). Time of day determines postexercise metabolism in mouse adipose tissue. Proceedings of the National Academy of Sciences of the United States of America, 120(8), e2218510120.

Publication 2023

NanoDrop One Agilent RNA 600 nano kit Bioanalyzer instrument TruSeq Stranded Total RNA with Ribo-Zero Gold protocol Qubit dsDNA HS assay kit NextSeq500

Assay Base pair Dna chip Dsdna Gold Sensitivity

Corresponding Organization :

Other organizations : Karolinska Institutet, Novo Nordisk Foundation, University of Copenhagen

Top 5 similar protocols

Variable analysis

independent variables

RNA concentration and purity assessed by absorbance at 260 and 280 nm using NanoDrop One
RNA quality checked using the Agilent RNA 600 nano kit and Bioanalyzer instrument
RNA aliquots (1,000 ng) analyzed using the Illumina TruSeq Stranded Total RNA with Ribo-Zero Gold protocol
Libraries cleaned and validated for DNA concentration using the Qubit dsDNA HS assay kit
Libraries validated for base pair size and purity using the Agilent High Sensitivity DNA chip and Bioanalyzer instrument
Libraries subjected to 38-bp paired-end sequencing on a NextSeq500

dependent variables

RNA concentration and purity
RNA quality
DNA concentration
Base pair size and purity of libraries

control variables

RNA concentration (1,000 ng aliquots)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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