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Trueseq stranded mrna library preparation kit

Manufactured by Illumina
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The TruSeq Stranded mRNA Library Preparation kit is a laboratory equipment product designed for preparing mRNA-derived sequencing libraries. The kit enables the conversion of mRNA into a library of cDNA fragments, which can then be sequenced using next-generation sequencing platforms. The core function of the kit is to provide a standardized and efficient workflow for preparing mRNA sequencing libraries.

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Lab products found in correlation

Truseq rna sample preparation kit v2, by Illumina (2 mentions) Hiseq 2000 v3 kits, by Illumina (2 mentions) Hiseq 2500, by Illumina (1 mentions) Qiaquick gel extraction kit, by Qiagen (1 mentions) Bigdye terminator v3.1 cycle sequencing kit, by Thermo Fisher Scientific (1 mentions) Abi prism 3730 genetic analyser, by Thermo Fisher Scientific (1 mentions) Novaseq 6000 sp flow cell, by Illumina (1 mentions) Spectrum plant total rna kit, by Merck Group (1 mentions) Dnase10, by Merck Group (1 mentions) Agilent 2200 tapestation system, by Agilent Technologies (1 mentions) Nextseq 500 sequencing platform, by Illumina (1 mentions)

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TrueSeq kits (3 citations)

5 protocols using trueseq stranded mrna library preparation kit

1

Sequencing of Plant Virus Genomes

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Purified dsRNAs were used for the preparation of a cDNA library using the True Seq Stranded mRNA Library preparation kit (Illumina) and sequenced in a multiplexed run of 100 bp single reads using a HiSeq2500, together with other banks prepared from unrelated plant samples. Both library construction and sequencing were performed by SEQMe s.r.o. (Czech Republic).
For targeted Sanger sequencing of coat protein and RdRp genes or RACE-PCR fragments (at least five clones) all nucleic acids were isolated using the QIAquick Gel Extraction kit (Qiagen). Isolated DNA was sequenced using the BigDye Terminator v3.1 Cycle Sequencing kit (Applied Biosystems) and an ABI PRISM 3730 Genetic analyser.
Šafářová D., Vavroušková K., Candresse T, & Navrátil M. (2018). Molecular characterization of a novel Aureusvirus infecting elderberry (Sambucus nigra L.). PLoS ONE, 13(8), e0200506.
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2

Illumina RNA-Seq Library Preparation

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mRNA in total RNA was isolated and converted into non-stranded or stranded libraries. Non-stranded libraries were produced manually using reagents provided in the Illumina TruSeq RNA Sample Preparation Kit v2 in accordance with manufacturer’s recommendations. The protocol was modified to produce size-selected libraries by modifying the fragmentation conditions and using a Caliper LabChip XT instrument. Stranded libraries were prepared using a NeoPrep Library Prep System and the reagents provided in the Illumina TrueSeq Stranded mRNA Library Preparation kit. The stranded library prep workflow is similar to the non-stranded workflow, except that it involves additional ribosomal reduction chemistry to maximise the percentage of uniquely mapped reads. Following purification, the RNA was fragmented and synthesised into cDNA using a reverse transcriptase process. The products were then enriched with PCR (maximum 10 cycles) to create the final cDNA library. Enriched libraries were subjected to 75 base paired-end sequencing using Illumina HiSeq 2000 v3 kits following manufacturer’s instructions.
Kilpinen H., Goncalves A., Leha A., Afzal V., Alasoo K., Ashford S., Bala S., Bensaddek D., Casale F.P., Culley O.J., Danecek P., Faulconbridge A., Harrison P.W., Kathuria A., McCarthy D., McCarthy S.A., Meleckyte R., Memari Y., Moens N., Soares F., Mann A., Streeter I., Agu C.A., Alderton A., Nelson R., Harper S., Patel M., White A., Patel S.R., Clarke L., Halai R., Kirton C.M., Kolb-Kokocinski A., Beales P., Birney E., Danovi D., Lamond A.I., Ouwehand W.H., Vallier L., Watt F.M., Durbin R., Stegle O, & Gaffney D.J. (2017). Common genetic variation drives molecular heterogeneity in human iPSCs. Nature, 546(7658), 370-375.
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3

Illumina RNA-Seq Library Preparation

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mRNA in total RNA was isolated and converted into non-stranded or stranded libraries. Non-stranded libraries were produced manually using reagents provided in the Illumina TruSeq RNA Sample Preparation Kit v2 in accordance with manufacturer’s recommendations. The protocol was modified to produce size-selected libraries by modifying the fragmentation conditions and using a Caliper LabChip XT instrument. Stranded libraries were prepared using a NeoPrep Library Prep System and the reagents provided in the Illumina TrueSeq Stranded mRNA Library Preparation kit. The stranded library prep workflow is similar to the non-stranded workflow, except that it involves additional ribosomal reduction chemistry to maximise the percentage of uniquely mapped reads. Following purification, the RNA was fragmented and synthesised into cDNA using a reverse transcriptase process. The products were then enriched with PCR (maximum 10 cycles) to create the final cDNA library. Enriched libraries were subjected to 75 base paired-end sequencing using Illumina HiSeq 2000 v3 kits following manufacturer’s instructions.
Kilpinen H., Goncalves A., Leha A., Afzal V., Alasoo K., Ashford S., Bala S., Bensaddek D., Casale F.P., Culley O.J., Danecek P., Faulconbridge A., Harrison P.W., Kathuria A., McCarthy D., McCarthy S.A., Meleckyte R., Memari Y., Moens N., Soares F., Mann A., Streeter I., Agu C.A., Alderton A., Nelson R., Harper S., Patel M., White A., Patel S.R., Clarke L., Halai R., Kirton C.M., Kolb-Kokocinski A., Beales P., Birney E., Danovi D., Lamond A.I., Ouwehand W.H., Vallier L., Watt F.M., Durbin R., Stegle O, & Gaffney D.J. (2017). Common genetic variation drives molecular heterogeneity in human iPSCs. Nature, 546(7658), 370-375.
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4

Plant Total RNA Extraction and Sequencing

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Whole-thorax samples were ground into a fine powder using a TissueLyser and used as input for the Spectrum Plant Total RNA Kit (STRN50, Sigma Aldrich), including DNase I treatment (DNASE10, Sigma Aldrich). Library preparation and sequencing were performed by SciLife Lab at the Uppsala Genome Centre. Sequencing libraries were prepared from 300 ng of RNA, using the TrueSeq stranded mRNA library preparation kit (20020595, Illumina Inc.) including polyA selection and unique dual indexing (20022371, Illumina Inc.), according to the manufacturer’s protocol. Sequencing was performed on the Illumina NovaSeq 6000 SP flowcell with paired-end reads of 150 bp.
Willink B., Tunström K., Nilén S., Chikhi R., Lemane T., Takahashi M., Takahashi Y., Svensson E.I, & Wheat C.W. (2023). The genomics and evolution of inter-sexual mimicry and female-limited polymorphisms in damselflies. Nature Ecology & Evolution, 8(1), 83-97.
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5

Stranded mRNA Sequencing Protocol

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The cDNA libraries were constructed using a TrueSeq Stranded mRNA library preparation kit (Illumina Inc., San Diego, CA, USA) according to the manufacturer’s instructions. Briefly, mRNA was separated from 300 to 500 ng of total RNA from each trachea using Oligo dT magnetic beads. First strand cDNA was synthesized from purified, fragmented mRNA using reverse transcriptase and random hexamer primers. The second strand was then synthesized using dUTP, dATP, dCTP, dGTP, DNA polymerase and RNase, and amplified by PCR. The end products were purified after end-repair and ligation of adapters, and the fragment size (~260 bp) and the quality of the cDNA libraries were confirmed using the Agilent 2200 TapeStation system. The libraries were normalized to 1 nM and then pooled, denatured and sequenced on a NextSeq500 sequencing platform (Illumina) to obtain 150 bp paired-end reads.
Kulappu Arachchige S.N., Young N.D., Kanci Condello A., Omotainse O.S., Noormohammadi A.H., Wawegama N.K, & Browning G.F. (2021). Transcriptomic Analysis of Long-Term Protective Immunity Induced by Vaccination With Mycoplasma gallisepticum Strain ts-304. Frontiers in Immunology, 11, 628804.
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