Truseq rna seq library prep kit v2

Manufactured by Illumina

Sourced in United States, China

The TruSeq RNA-Seq Library Prep Kit-v2 is a laboratory equipment used for preparing RNA sequencing libraries. It provides a streamlined workflow for converting RNA samples into libraries suitable for sequencing on Illumina platforms.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (1 mentions) Hiseq 2500 sequencing system, by Illumina (1 mentions) Rna 6000 nano labchip, by Agilent Technologies (1 mentions) 2100 bioanalyzer, by Agilent Technologies (1 mentions) Paxgene blood mirna kit, by Qiagen (1 mentions) Hiseq 2500 platform, by Illumina (1 mentions) Nanodrop nd 2000 ultra micro spectrophotometer, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Hiseq 2000, by Illumina (1 mentions)

4 protocols using truseq rna seq library prep kit v2

RNA-seq Analysis of Differentially Expressed Genes

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Total RNA was extracted with TRIzol (Life Technologies) according to the manufacturer’s instructions¹⁵. RNA quality and quantity were assessed by using a Bioanalyzer 2100 with RNA 6000 Nano Labchips (Agilent Technologies, Dublin, Ireland). The preparation of RNA libraries using the Truseq RNA-Seq Library Prep Kit-v2 and paired-end sequencing using the HiSeq 2500 sequencing system (Illumina, San Diego, CA, USA) were performed by Macrogen (Seoul, Korea). The RNA sequencing data were deposited in the Gene Expression Omnibus GEO database under accession number GSE139887.
Differentially expressed genes (DEGs) were filtered as previously reported¹⁴, except that the genes showing fragments per kilobase of transcript per million mapped reads (FPKM) values below 10 were excluded from the subsequent analysis. Ingenuity pathway analysis (IPA^®, QIAGEN, Redwood City, CA, USA) was also performed to identify key biological functions based on the curated diseases and functional ontologies in the IPA knowledge database, as previously reported¹⁴. Multi-Experiment Viewer 4.9.0 (mev.tm4.org) was used for the graphic representation of the values.

Poojan S., Bae S.H., Min J.W., Lee E.Y., Song Y., Kim H.Y., Sim H.W., Kang E.K., Kim Y.H., Lee H.O., Hong Y., Park W.Y., Jang H, & Hong K.M. (2020). Cancer cells undergoing epigenetic transition show short-term resistance and are transformed into cells with medium-term resistance by drug treatment. Experimental & Molecular Medicine, 52(7), 1102-1115.

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RNA Extraction and Sequencing from Blood

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Blood (2.5 mL) was taken from each patient into a PAXgene Blood RNA Tube, containing a patented RNA stabilizer reagent composition. RNA was automated isolated in QIAGEN’s QIAcube using the PAXgene Blood miRNA Kit from QIAGEN PreAnalytiX. RNA sequencing libraries were prepared according to the Illumina TruSeq® messenger (mRNA) sequencing protocol (TruSeq® RNA Seq Library Prep Kit v2). The resulting libraries were sequenced on the NovaSeq 6000 (2 × 50 bp, S2 chemistry).

Bernardes J.P., Mishra N., Tran F., Bahmer T., Best L., Blase J.I., Bordoni D., Franzenburg J., Geisen U., Josephs-Spaulding J., Köhler P., Künstner A., Rosati E., Aschenbrenner A.C., Bacher P., Baran N., Boysen T., Brandt B., Bruse N., Dörr J., Dräger A., Elke G., Ellinghaus D., Fischer J., Forster M., Franke A., Franzenburg S., Frey N., Friedrichs A., Fuß J., Glück A., Hamm J., Hinrichsen F., Hoeppner M.P., Imm S., Junker R., Kaiser S., Kan Y.H., Knoll R., Lange C., Laue G., Lier C., Lindner M., Marinos G., Markewitz R., Nattermann J., Noth R., Pickkers P., Rabe K.F., Renz A., Röcken C., Rupp J., Schaffarzyk A., Scheffold A., Schulte-Schrepping J., Schunk D., Skowasch D., Ulas T., Wandinger K.P., Wittig M., Zimmermann J., Busch H., Hoyer B.F., Kaleta C., Heyckendorf J., Kox M., Rybniker J., Schreiber S., Schultze J.L, & Rosenstiel P. (2020). Longitudinal Multi-omics Analyses Identify Responses of Megakaryocytes, Erythroid Cells, and Plasmablasts as Hallmarks of Severe COVID-19. Immunity, 53(6), 1296-1314.e9.

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RNA-Seq Analysis of Duck Adipocytes

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Duck pre-adipocytes and those that had undergone differentiation for 3 days (n = 3) were collected for RNA-sequencing analyses of ribosomal RNA-depleted total RNA analyses. Thereafter, RNA libraries were constructed using the Truseq RNA-Seq Library Prep Kit v.2 (Illumina) following the manufacturer’s instructions. Libraries were sequenced using the Illumina HiSeq 2500 platform by paired-end sequencing, and this was completed by Novogene Biotechnology Co., Ltd. (Beijing, China).

Wang L., Liang W., Wang S., Wang Z., Bai H., Jiang Y., Bi Y., Chen G, & Chang G. (2020). Circular RNA expression profiling reveals that circ-PLXNA1 functions in duck adipocyte differentiation. PLoS ONE, 15(7), e0236069.

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Transcriptomic Analysis of Abdominal Fat Tissue

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Total RNA was extracted from the abdominal fat tissue samples using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). RNA concentration and quality were measured at OD 260/280 using the Nanodrop ND-2000 ultra-micro spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA). OD 260/280 in the range of 2.0–2.2 and RNA concentration around 500 ng/μL. The integrity of the RNA was measured by analyzing 2 µL of the total RNA on a 1% agarose gel. RNA libraries were then constructed using the TruSeq RNA-Seq Library Prep Kit v.2 (Illumina, Shanghai, China) following the manufacturer’s instructions. Subsequently, libraries were constructed and sequenced using a HiSeqTM 2000 instrument (Illumina), completed by Gene Denovo Biotechnology Co., Ltd. (Guangzhou, China).
To ensure high-quality clean reads, FASTP was used for quality control. The resulting high-quality clean reads were compared with a ribosomal database using Bowtie2 [15 (link)]. This comparison allowed for calculating the ratio of high-quality clean data to total RNA. The reads derived from the filtered ribosomes were aligned to the reference genome using TopHat2 [16 (link)]. Unmapped reads were extracted from the alignment results, and both ends of each unmapped read (default 20 bp) were intercepted to obtain the anchor reads. The anchor reads were then aligned to the reference genome.

Yang Y., Yang C., Zhuang Z., Mao J., Chen A., Zhou T., Bai H., Jiang Y., Chang G, & Wang Z. (2024). RNA-Seq Analysis Revealed circRNAs and Genes Associated with Abdominal Fat Deposition in Ducks. Animals : an Open Access Journal from MDPI, 14(2), 260.

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