Truseq stranded mrna lt prep kit

Manufactured by Illumina

Sourced in United States

The TruSeq Stranded mRNA LT Prep Kit is a laboratory equipment product designed for mRNA sample preparation prior to next-generation sequencing. It provides a streamlined workflow for isolating, fragmenting, and converting mRNA into a library of cDNA fragments with adapter sequences attached to both ends.

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Lab products found in correlation

Hiseq 2500, by Illumina (2 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Nextseq 500, by Illumina (1 mentions) Click it nascent rna capture kit, by Thermo Fisher Scientific (1 mentions) Ribo zero gold rrna removal kit, by Illumina (1 mentions) Microelute total rna kit, by Omega Bio-Tek (1 mentions)

Spelling variants of this product

TruSeq Stranded mRNA LT Sample Prep Kit (534 citations) TruSeq Stranded mRNA (115 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 Library Prep Kit LT (6 citations) TruSeq Stranded mRNA LT Sample Prep Kit—Set A (3 citations) RNA TruSeq Stranded mRNA LT sample prep kit (2 citations)

4 protocols using truseq stranded mrna lt prep kit

RNA-seq Analysis of Differential Gene Expression

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RNA-seq and bioinformatics analysis were performed similar to our previous study [75 (link)]. Briefly, RNA was isolated using Trizol (Invitrogen, Carlsbad, CA, USA), and cDNA libraries were prepared using the TruSeq Stranded mRNA LT Prep Kit (Illumina). Libraries were sequenced on a HiSeq 2500 instrument (Illumina) at the MGH Next Generation Sequencing Core Facility, using paired-end 50-bp sequencing. Sequencing reads were mapped by Novogene (Beijing, China). Read counts over transcripts were calculated using HTseq, followed by differential expression analysis using EdgeR. Genes were classified as differentially expressed based on the cutoffs of fold change (FC) > 1.6, false discovery rate (FDR) < 0.1, and p < 0.005. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of differentially expressed genes (DEGs) were performed using the R package (v 3.5.1).

Zhang W., Ding L., Chen H., Zhang M., Ma R., Zheng S., Gong J., Zhang Z., Xu H., Xu P, & Zhang Y. (2023). Cntnap4 partial deficiency exacerbates α-synuclein pathology through astrocyte–microglia C3-C3aR pathway. Cell Death & Disease, 14(4), 285.

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Transcriptional Profiling of Mouse and Human Hematopoietic Stem Cells

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Total RNA was isolated from mouse HSCs and human HSPCs using PicoPure RNA isolation kit (KIT0204). RNA from mESC and hESC was isolated using the Qiagen mini kit. 100 ng of total RNA was used was used as input for the Illumina TruSeq Stranded mRNA LT Prep Kit (20020594). Libraries were made following Illumina’s recommended protocol. Amplified libraries were purified and quantified using the KAPA quantification kit. RNA sequencing libraries were sequenced on an Illumina NextSeq 500 instrument (paired-end 75bp).

Ramabadran R., Wang J.H., Reyes J.M., Guzman A.G., Gupta S., Rosas C., Brunetti L., Gundry M.C., Tovy A., Long H., Gu T., Cullen S.M., Tyagi S., Rux D., Kim J.J., Kornblau S.M., Kyba M., Stossi F., Rau R.E., Takahashi K., Westbrook T.F, & Goodell M.A. (2023). DNMT3A-coordinated splicing governs the stem state switch toward differentiation in embryonic and hematopoietic stem cells. Nature cell biology, 25(4), 528-539.

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Nascent RNA Isolation from Mouse and Human ESCs

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Mouse and Human ESCs were grown in 6-well plates. 0.2uM of EU was added for 6 hours. Nascent RNA was labelled and isolated using the Click-IT Nascent RNA Capture kit (C10365) kit from Invitrogen. Briefly, RNA was extracted using Trizol and total RNA was ribo-depleted with Illumina’s Ribo-Zero Gold rRNA Removal Kit (MRZG126) as per manufacturer’s recommendations. 2ug of Ribo-depleted Total RNA was subjected to further selection by Poly A purification beads provided in Illumina’s TruSeq Stranded mRNA LT Prep kit. Supernatant from incubation with beads was used as nascent RNA (Poly A⁻ and Ribo-depleted).

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Transcriptomic Profiling of Microglia in 5xFAD Mice

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Microglia were FACS sorted from brains of 5-month-old animals as described above. Microglia cells were isolated from 12 5xFAD mice (6M/6F) and 12 Il3^−/−5xFAD mice (6M/6F). Within each genotype samples from 2M and 2F were pooled generating 3 samples from 5xFAD mice and 3 from Il3^−/−5xFAD mice from which RNA was isolatedd the RNA-seq performed. RNA was isolated using E.Z.N.A micro elute total RNA kit according to the manufacturer’s instructions (Omega Biotek). cDNA libraries were prepared using the TruSeq Stranded mRNA LT Prep Kit (Illumina). Libraries were sequenced on a HiSeq 2500 instrument (Illumina) at the MGH Next Generation Sequencing Core Facility, using paired-end 50 bp sequencing. Sequencing reads were mapped in a splice-aware fashion to the Ensembl annotation of the mouse GRCm37/mm9 transcriptome. Read counts over transcripts were calculated using HTseq followed by differential expression analysis using EdgeR. Genes were classified as differentially expressed based on the cutoffs of fold change (FC)>1.6, false discovery rate (FDR)<0.1, and p<0.005.

McAlpine C.S., Park J., Griciuc A., Kim E., Choi S.H., Iwamoto Y., Kiss M.G., Christie K.A., Vinegoni C., Poller W.C., Mindur J.E., Chan C.T., He S., Janssen H., Wong L.P., Downey J., Singh S., Anzai A., Kahles F., Jorfi M., Feruglio P.F., Sadreyev R.I., Weissleder R., Kleinstiver B.P., Nahrendorf M., Tanzi R.E, & Swirski F.K. (2021). Astrocyte-derived interleukin-3 reprograms microglia and limits Alzheimer’s disease. Nature, 595(7869), 701-706.

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