Dynabeads mrna kit

Manufactured by Thermo Fisher Scientific

The Dynabeads mRNA kit is a laboratory product designed for the isolation and purification of messenger RNA (mRNA) from biological samples. It utilizes magnetic beads coated with oligo(dT) to selectively capture and separate mRNA molecules from total RNA extracts.

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Lab products found in correlation

Scriptseq rna seq library preparation kit, by Illumina (1 mentions) Rnalater stabilization reagent, by Qiagen (1 mentions) Qiaxcel, by Qiagen (1 mentions) Affinity script enzyme, by Agilent Technologies (1 mentions) T4 ligase, by New England Biolabs (1 mentions) Hiseq 2500, by Illumina (1 mentions)

Close spelling variants of this product

Dynabead direct mRNA extraction kit (2 citations) Dynabead™ mRNA DIRECT™ Micro kit (5 citations) Dynabead mRNA DIRECT kit (9 citations) Dynabead mRNA DIRECT Micro Purification Kit (2 citations) Dynabead mRNA Purification Kit (11 citations) Dynabeads mRNA Purification Kit (522 citations) Ambion Dynabeads mRNA Direct Kit (2 citations) Dynabeads Oligo (dT)25 mRNA Purification Kit (2 citations) Dynabeads mRNA Purification Kit for mRNA Purification from Total RNA preps (3 citations) Ambion Dynabeads mRNA Purification Kit (2 citations)

2 protocols using dynabeads mrna kit

Spleen and Lymph Node RNA-Seq

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Spleen and cervical lymph nodes were dissected fresh into RNALater stabilization reagent (Qiagen) and stored at −20 °C. Total RNA was isolated using MagMAX™ Nucleic Acid Isolation Kit (Ambion) per the manufacturer’s instructions. RNA was quantified using UV spectrophotometer, and RNA integrity was evaluated by Qiaxcel (Qiagen). PolyA mRNA was purified from total RNA using Dynabeads mRNA kit (Invitrogen) and strand specific RNA-Seq libraries were prepared with the ScriptSeq RNA-seq Library Preparation kit (Illumina). RNA-Seq libraries were sequenced to a length of 33 bp using Hiseq 2000 NGS sequencer (Illumina). Gene expression levels were derived from raw sequencing reads using Nimbus2, an RNA-Seq analysis pipeline developed in house.

Atanasio A., Decman V., White D., Ramos M., Ikiz B., Lee H.C., Siao C.J., Brydges S., LaRosa E., Bai Y., Fury W., Burfeind P., Zamfirova R., Warshaw G., Orengo J., Oyejide A., Fralish M., Auerbach W., Poueymirou W., Freudenberg J., Gong G., Zambrowicz B., Valenzuela D., Yancopoulos G., Murphy A., Thurston G, & Lai K.M. (2016). C9orf72 ablation causes immune dysregulation characterized by leukocyte expansion, autoantibody production, and glomerulonephropathy in mice. Scientific Reports, 6, 23204.

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Strand-specific RNA-seq library preparation

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mRNA was prepared using Dynabeads mRNA kit (Invitrogen) and fragmented by heat. For cDNA synthesis, custom oligo-dT primers were used with a barcode and an adapter-linker sequence (CAG ACG TGT GCT CTT CCG ATC T—XXXXXXXX-T15) and with the Affinity Script enzyme (Agilent). After first strand synthesis, samples were pooled together based on ACTB qPCR values. RNA-DNA hybrids was degraded using consecutive acid-alkali treatment. A second sequencing linker (AGA TCG GAA GAG CGT CGT GTAG) was then ligated using T4 ligase (NEB). The mixture was then PCR enriched for 12 cycles and purified to yield final strand specific RNA-seq libraries as previously described^{55 (link)}. Libraries were sequenced using a HiSeq 2500 (Illumina) using 50bp × 25bp pair-end sequencing.

Chatterjee S., Luthra P., Esaulova E., Agapov E., Yen B.C., Borek D.M., Edwards M.R., Mittal A., Jordan D.S., Ramanan P., Moore M.L., Pappu R.V., Holtzman M.J., Artyomov M., Basler C.F., Amarasinghe G.K, & Leung D.W. (2017). Structural basis for human respiratory syncytial virus NS1-mediated modulation of host responses. Nature microbiology, 2, 17101.

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