Truseq stranded mrna library prep kit lt

Manufactured by Illumina

Sourced in United States

The TruSeq Stranded mRNA Library Prep Kit LT is a laboratory equipment product designed for the preparation of stranded mRNA libraries for RNA sequencing applications. The kit provides the necessary reagents and protocols to convert mRNA into a library of cDNA fragments with adapter sequences attached to both ends.

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Lab products found in correlation

Bcl2fastq v1, by Illumina (2 mentions) Bioanalyzer, by Agilent Technologies (2 mentions) Hiseq 2500 rapid run flow cell, by Illumina (2 mentions) Hiseq 2500 sequencer, by Illumina (1 mentions) Nextseq 550 sequencing platform, by Illumina (1 mentions) Direct zol rna microprep kit, by Zymo Research (1 mentions) Ercc rna spike in mix, by Thermo Fisher Scientific (1 mentions) Rneasy plant mini kit, by Qiagen (1 mentions) Rna nano 6000 assay kit, by Agilent Technologies (1 mentions) Nextseq 500 system, by Illumina (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions) Agilent bioanalyzer dna 1000 kit, by Agilent Technologies (1 mentions) Real time analysis v1, by Illumina (1 mentions) Nextseq 500, by Illumina (1 mentions) 2100 bioanalyzer, by Agilent Technologies (1 mentions)

Spelling variants of this product

TruSeq Stranded mRNA (115 citations) TruSeq Stranded mRNA Library Prep (98 citations) TruSeq Stranded mRNA LT Kit (62 citations) TruSeq kits (44 citations) Stranded mRNA Prep (27 citations) TruSeq Stranded mRNA LT (9 citations) MRNA TruSeq kit (7 citations) TruSeq Stranded mRNA LT Prep Kit (4 citations)

6 protocols using truseq stranded mrna library prep kit lt

mRNA Sequencing Library Preparation

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One microgram of total RNA per each time point was used as the input to an mRNA capture with oligo-dT coated magnetic beads. The mRNA was fragmented, and then a random-primed cDNA synthesis was performed. The resulting double-strand cDNA was used as the input to a standard Illumina library prep (TruSeq Stranded mRNA LT library prep kit, Illumina) with end-repair, adapter ligation and PCR amplification being performed to generate the library. The indexed mRNAseq libraries were quantitated by qPCR, pooled with equimolar amounts and sequenced on an Illumina HiSeq 2500 sequencer for a 2 x125 cycle run.

Greer Y.E., Porat-Shliom N., Nagashima K., Stuelten C., Crooks D., Koparde V.N., Gilbert S.F., Islam C., Ubaldini A., Ji Y., Gattinoni L., Soheilian F., Wang X., Hafner M., Shetty J., Tran B., Jailwala P., Cam M., Lang M., Voeller D., Reinhold W.C., Rajapakse V., Pommier Y., Weigert R., Linehan W.M, & Lipkowitz S. (2018). ONC201 kills breast cancer cells in vitro by targeting mitochondria. Oncotarget, 9(26), 18454-18479.

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Transcriptome Analysis of Drosophila Embryos

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Embryos from the Df(2L)His C FRT40A/CyO Dfd-GFP; 6xHisGU and Df(2L)His C FRT40A/CyO Dfd-GFP; 6xHisGU H3K14R strains were collected on apple juice agar plates supplemented with yeast for one hour and aged an additional 12 hours. After dechorionation in bleach, they were hand-sorted according to GFP expression, collected in 300 uL TRIzol LS Reagent, snap-frozen in liquid N 2 and stored at À80 C until further use. On the day of the extraction, the embryos were homogenized in TRIzol LS Reagent using a 1.5 mL tube pestle and the total RNA was purified using the Direct-zol RNA MicroPrep kit (R2060, Zymo Research). Twenty embryos were used in each replicate. Libraries were prepared using the TruSeq Stranded mRNA LT Library Prep Kit (Illumina), spiked-in with ERCC RNA Spike-In Mix (Invitrogen) and single-end (1x75 bp) sequenced in the NextSeq 550 Sequencing platform (Illumina) at BEA core facility, Stockholm.

Regadas I., Dahlberg O., Vaid R., Ho O., Belikov S., Dixit G., Deindl S., Wen J, & Mannervik M. (2021). A unique histone 3 lysine 14 chromatin signature underlies tissue-specific gene regulation. Molecular cell, 81(8).

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RNA Extraction and Sequencing from Plant Roots

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We extracted total RNA from the plant roots using Qiagen RNeasy® Plant Mini Kit (Cat #74904, California, USA) as described in Hiltenbrand et al. (2016) (link). For hormone-treated plants, RNA was extracted only from plant roots that had NLS. We performed RNA quantification, library preparation, and RNA sequencing at the Research Technology Support Facility (RTSF), Michigan State University, East Lansing, MI. Before library preparation and sequencing, the RNA integrity was checked using a Bioanalyzer (Agilent Technologies). We used the Illumina TruSeq Stranded mRNA Library Prep Kit LT to prepare multiplex sequencing libraries. These six libraries were combined into a single pool, loaded on one lane of an Illumina HiSeq 2500 Rapid Run flow cell (v2), and sequenced in a PE100 format with HiSeq Rapid SBS reagents. We used Illumina Real Time Analysis (RTA) v1.18.64 for base calling and used Illumina Bcl2fastq v1.8.4 to de-multiplex the output of RTA and convert to Fastq format.

Thomas J., Bowman M.J., Vega A., Kim H.R, & Mukherjee A. (2018). Comparative transcriptome analysis provides key insights into gene expression pattern during the formation of nodule-like structures in Brachypodium. Functional & integrative genomics, 18(3), 315-326.

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Macaque Heart RNA-Seq Analysis

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Macaque hearts were used to extract total RNA, which was then tested using an RNA Nano 6000 Assay Kit and an Agilent 2100 bioanalyzer (Agilent Technologies, CA, USA). Library creation was done using a TruSeq Stranded mRNA Library Prep Kit LT (Illumina, San Diego, CA, USA) in accordance with the manufacturer's instructions for library preparation and sequencing. Using the Agilent Bioanalyzer DNA‐1000 Kit (Agilent Technologies), library quality and size distribution were assessed. The libraries' average sizes were roughly 260 bp, as advised by Illumina. With the use of the Bio‐Rad CFX 96 KIT IQ SYBR GRN, quantitative PCR was carried out to pool equivalent numbers of libraries with various adaptor indices. The NextSeq 500 system (Illumina) was used for all sequencing, and a NextSeq500 mid‐output v2 paired‐end sequencing kit with 150 cycles was used (Illumina).

Li H., Wang T., Feng Y., Sun K., Huang G., Cao Y, & Xu A. (2023). Optimal transplantation strategy using human induced pluripotent stem cell‐derived cardiomyocytes for acute myocardial infarction in nonhuman primates. MedComm, 4(3), e289.

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Illumina-based RNA Sequencing Workflow

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RNA sequencing including RNA quantification and library preparation was performed at the Research Technology Support Facility (RTSF), Michigan State University, East Lansing, MI, USA. The RNA was checked for integrity before library preparation and sequencing using a Bioanalyzer (Agilent Technologies). Briefly, multiplex sequencing libraries were prepared using the Illumina TruSeq Stranded mRNA Library Prep Kit LT. After QC, all six libraries (three treatment and three controls) were combined into a single pool. This pool was loaded on one lane of an Illumina HiSeq 2500 Rapid Run flow cell (v1). Sequencing was performed in a PE100 format with Rapid SBS reagents. Base calling was done by Illumina Real Time Analysis (RTA) v1.17.21.3 and output of RTA was de-multiplexed and converted to Fastq format with Illumina Bcl2fastq v1.8.4.

Hiltenbrand R., Thomas J., McCarthy H., Dykema K.J., Spurr A., Newhart H., Winn M.E, & Mukherjee A. (2016). A Developmental and Molecular View of Formation of Auxin-Induced Nodule-Like Structures in Land Plants. Frontiers in Plant Science, 7, 1692.

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RNA Sequencing Library Preparation

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Total RNA was extracted from two biological replicates of each sample using Trizol method with DNase treatment, and RNA quality was assessed using 2100 Bioanalyzer (Agilent Genomics). Samples with RIN ≥ 9 proceeded to library preparation using TruSeq stranded mRNA Library Prep Kit LT (Illumina) according to the manufacturer’s protocol. Library concentration was determined by qRT-PCR (KAPA Library Quantification Kit) to pool equal amounts of libraries with different adapter indexes. Sequencing was performed using NextSeq500 (Illumina). Data were analyzed as described in Extended Experimental Procedures.

Chmielowiec J., Szlachcic W.J., Yang D., Scavuzzo M.A., Wamble K., Sarrion-Perdigones A., Sabek O.M., Venken K.J, & Borowiak M. (2022). Human pancreatic microenvironment promotes β-cell differentiation via non-canonical WNT5A/JNK and BMP signaling. Nature Communications, 13, 1952.

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