Cells were sorted (BD DiVa) based on tdTomato, CD24, EpCam, and CD45 as indicated in the figure legend then RNA was prepped from at least (Mock: 78,000, 2 DPI: 4,100, 14 DPI SC: 44,000, 14 DPI: 100,000) cells using RNeasy Plus Mini kit (74104). Samples were then prepped using NEBNext Poly(A) messenger RNA (mRNA) Magnetic Isolation Module (NEB #E74905). NEBNext Ultra II RNA Library Prep Kit (E7770S) was used to prep libraries with NEBNext Multiplex Oligos for Illumina (E7335S, E7500S) for adaptor ligation and enrichment of libraries. Samples were analyzed on two lanes of Illumina HiSeq 2500 in rapid run mode. The raw and normalized data is accessible at NCBI GEO under the accession number GSE116032. The reads were mapped to the mouse transcriptome (mm10) and the B/Mal/04 genome using Bowtie, and samples were run on Basespace (Illumina). Finally reads were imported into DeSeq2 for analysis65 (link).
Nebnext poly a messenger rna mrna magnetic isolation module
Manufactured by New England Biolabs
The NEBNext Poly(A) messenger RNA (mRNA) Magnetic Isolation Module is a laboratory product designed to isolate and purify polyadenylated mRNA from total RNA samples. The module utilizes magnetic beads coated with oligo(dT) sequences to selectively capture and separate mRNA molecules from other RNA species.
Automatically generated - may contain errors
Lab products found in correlation
Nebnext multiplex oligos for, by Illumina (2 mentions)
Novaseq 6000, by Illumina (2 mentions)
Basespace, by Illumina (1 mentions)
Nebnext ultra 2 rna library prep kit, by New England Biolabs (1 mentions)
Hiseq 2500, by Illumina (1 mentions)
Monarch total rna miniprep kit, by New England Biolabs (1 mentions)
Total rneasy mini kit, by Qiagen (1 mentions)
Nebnext ultra 2 directional rna library prep kit for, by Illumina (1 mentions)
3 protocols using nebnext poly a messenger rna mrna magnetic isolation module
1
Single-cell RNA-seq of influenza infection
2
Transcriptome Analysis of SARS-CoV-2 Infection in Huh7 Cells
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ZBTB7A-Huh7 cells and control cells were seeded into 6-well plates and mock-infected or infected with HCoV-229E at 1 MOI. Each group contained 4 replicates. The cells were collected 24 hpi and total RNA was extracted with Monarch Total RNA Miniprep Kit (NEB, T2010S). Samples were prepared with a NEBNext Poly(A) messenger RNA (mRNA) Magnetic Isolation Module (NEB #E74905). The NEBNext Ultra II RNA Library Prep Kit (E7770S) was used to prepare libraries with NEBNext Multiplex Oligos for Illumina (E7335S, E7500S). Samples were then sequenced on an Illumina NovaSeq 6000 and then raw reads were mapped to the human reference genome. After normalization, average read values were compared across samples. The data were further sorted based on the fold change and p-values. The upregulated and downregulated genes were categorized by gene ontology (pantherdb.org ). The raw sequencing data are available at NCBI GEO under accession number: GSE197644 .
3
RNA-seq Library Preparation Protocol
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RNA was isolated using Total RNeasy Mini Kit (QIAGEN Inc.). RNA‐seq libraries were prepared using NEBNext Poly(A) messenger RNA (mRNA) Magnetic Isolation Module (NEB #E7490) and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760). RNA‐seq libraries were validated by MedGenome (San Francisco, CA) before being sequenced on a Nova‐Seq 6000 (Illumina).
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