MDM were generated as described and stimulated with 25 ng/ml of the indicated IFNs. Total RNA was isolated 18 h following stimulation using RNeasy minikit (Qiagen). Intact poly(A) RNA was purified from total RNA samples (100 to 500 ng) with oligo(dT) magnetic beads, and stranded mRNA sequencing libraries were prepared as described using the Illumina TruSeq Stranded mRNA Library Preparation kit (catalog no. RS-122-2101 and RS-122-2102). Purified libraries were qualified on an Agilent Technologies 2200 TapeStation using a D1000 ScreenTape assay (catalog no. 5067-5582 and 5067-5583). The molarity of adapter-modified molecules was defined by quantitative PCR using the Kapa Biosystems Kapa Library Quant kit (catalog no. KK4824). Individual libraries were normalized to 10 nM, and equal volumes were pooled in preparation for Illumina sequence analysis. Sequencing libraries (25 pM) were chemically denatured and applied to an Illumina HiSeq v4 single-read flow cell using an Illumina cBot system. Hybridized molecules were clonally amplified and annealed to sequencing primers with reagents from an Illumina HiSeq SR cluster kit v4-cBot (catalog no. GD-401-4001). Following transfer of the flow cell to an Illumina HiSeq 2500 instrument (HCSv2.2.38 and RTA v1.18.61), a 50-cycle single-read sequence run was performed using HiSeq SBS kit v4 sequencing reagents (catalog no. FC-401-4002).
Hiseq sbs kit v4 sequencing reagents
Manufactured by Illumina
Sourced in United States
The HiSeq SBS Kit v4 is a set of sequencing reagents designed for use with Illumina's HiSeq sequencing systems. The kit provides the necessary reagents for performing sequencing-by-synthesis (SBS) reactions, which is the core functionality of the HiSeq platforms. The reagents enable the sequencing of DNA samples on the HiSeq instruments.
Automatically generated - may contain errors
Lab products found in correlation
Truseq stranded mrna library preparation kit, by Illumina (3 mentions)
Hiseq sr cluster kit v4 cbot, by Illumina (3 mentions)
Hiseq v4 single read flow cell, by Illumina (3 mentions)
Rneasy mini kit, by Qiagen (3 mentions)
Truseq stranded mrna sample preparation kit, by Illumina (3 mentions)
Hiseq 2500 instrument, by Illumina (2 mentions)
Hiseq pe cluster kit v4 cbot, by Illumina (2 mentions)
Hiseq instrument, by Illumina (2 mentions)
Hiseq 2500, by Illumina (2 mentions)
Cbot system, by Illumina (1 mentions)
Kapa library quant kit, by Roche (1 mentions)
D1000 screentape assay, by Agilent Technologies (1 mentions)
Tapestation 2200, by Agilent Technologies (1 mentions)
Hiseqv4, by Illumina (1 mentions)
Hiseq 2500 platform, by Illumina (1 mentions)
Hiseq 2500 sequencer, by Illumina (1 mentions)
Direct zol rna extraction kit, by Zymo Research (1 mentions)
Artseq, by Illumina (1 mentions)
Artseq ribosome profiling kit, by Illumina (1 mentions)
Multiplex small rna library prep set, by New England Biolabs (1 mentions)
8 protocols using hiseq sbs kit v4 sequencing reagents
1
RNA-seq protocol for IFN-stimulated MDM
2
Metagenome Sequencing on Illumina HiSeq 2500
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Metagenome libraries were constructed with size-selected, sonicated DNA fragments of 500 to 700 bp with the NEBnext Ultra DNA II library kit for Illumina (E7645S), as previously described (94 (link)). Paired-end sequencing (2 × 125 bp) of metagenomic libraries was conducted at the University of Utah High-Throughput Genomics Core Facility at the Huntsman Cancer Institute with an Illumina HiSeq 2500 platform. Sequencing libraries (25 pM) were chemically denatured and applied to an Illumina HiSeq v4 paired end flow cell using an Illumina cBot. Hybridized molecules were clonally amplified and annealed to sequencing primers with reagents from an Illumina HiSeq PE Cluster Kit v4-cBot. Following transfer of the flowcell to an Illumina HiSeq 2500 instrument (HCS v2.2.38 and RTA v1.18.61), a 125 cycle paired-end sequence run was performed using HiSeq SBS Kit v4 sequencing reagents (FC-401-4003).
3
RNA Isolation and Sequencing of Mouse Tumors
4
Transcriptomic Analysis of Plant Responses
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Total RNA was prepared from leaf material from Tanglefoot enclosures with the DirectZol RNA extraction kit (Zymo Research, Irvine, CA, United States). Barcoded RNA-seq libraries were constructed at the High-Throughput Genomics Core Facility (University of Utah) using the Illumina (Illumina, San Diego, CA, United States) TruSeq Stranded mRNA Library Preparation Kit with poly(A) selection, and 125 bp paired-end reads were generated on an Illumina HiSeq 2500 sequencer with HiSeq SBS Kit v4 sequencing reagents. Briefly, four lanes were run in total for each of the barley and maize experiments, each consisting of 28 samples (four replicates each for the control, wounding at 2 and 24 h, T. urticae infestation at 2 and 24 h, and O. pratensis infestation at 2 and 24 h). Biological replicates were evenly distributed across the four sequencing lanes to reduce the possibility of confounding due to lane-level effects.
5
Interferon-Stimulated Transcriptome of HIV-Infected Monocyte-Derived Macrophages
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RNA sequencing data were generated by Szaniawski et al. [19 (link)]. The sequencing data are publicly available at the National Center of Biotechnology Information under accession no. GSE158434.
Briefly, CD14+ monocytes from healthy donors were cultured for 7 days to allow differentiation to MDM. MDM were stimulated in triplicate with 25 ng/mL of a single interferon (IFN-α, IFN-ε, IFN-γ or IFN-λ) or infected with 250 ng HIV-1-BAL-HSA virus. Total RNA was isolated 18 h following interferon stimulation or 6 h post infection using the RNeasy minikit (Qiagen, Hilden, Germany). Intact poly(A) RNA was purified and stranded mRNA sequencing libraries were prepared using the Illumina (San Diego, CA, USA) TruSeq Stranded mRNA Library Preparation kit (catalog No. RS-122-2101 and RS-122-2102). On an Illumina HiSeq 2500 instrument (HCSv2.2.38 and RTA v1.18.61), a 50-cycle single-read sequence run was performed using HiSeq SBS kit v4 sequencing reagents (catalog no. FC-401-4002).
Briefly, CD14+ monocytes from healthy donors were cultured for 7 days to allow differentiation to MDM. MDM were stimulated in triplicate with 25 ng/mL of a single interferon (IFN-α, IFN-ε, IFN-γ or IFN-λ) or infected with 250 ng HIV-1-BAL-HSA virus. Total RNA was isolated 18 h following interferon stimulation or 6 h post infection using the RNeasy minikit (Qiagen, Hilden, Germany). Intact poly(A) RNA was purified and stranded mRNA sequencing libraries were prepared using the Illumina (San Diego, CA, USA) TruSeq Stranded mRNA Library Preparation kit (catalog No. RS-122-2101 and RS-122-2102). On an Illumina HiSeq 2500 instrument (HCSv2.2.38 and RTA v1.18.61), a 50-cycle single-read sequence run was performed using HiSeq SBS kit v4 sequencing reagents (catalog no. FC-401-4002).
6
Ribosome Profiling of Platelets
7
RNA-Seq Analysis of DNA Methylation
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DNA methylation is an epigenetic mechanism that usually affects CpG islands and promoters regulating the expression of the genes. For this reason, a transcriptomic analysis by RNA-Seq was performed. Library construction was performed using an Illumina TruSeq Stranded mRNA Sample Preparation Kit (CAT. No. RS-122-2101, RS-122-2102). Following the transfer of the flow cell to an Illumina HiSeq instrument, a 101-cycle paired-end sequence run was performed using the HiSeq SBS Kit v4 sequencing reagents (FC-401-4002). A detailed description of the analysis is provided in the Additional file 1 . Sequencing data have been posted in the Gene Expression Omnibus (GEO) database under accession number GSE114575.
8
RNA-seq Library Preparation and Sequencing
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RNA was extracted from cells using RNeasy Kit from Qiagen. The RNA-seq libraries were generated using Illumina TruSeq Stranded mRNA sample preparation kit with oligo dT selection. Sequencing libraries (25 pM) were chemically denatured and applied to an Illumina HiSeq v4 paired end flow cell using an Illumina cBot. Hybridized molecules were clonally amplified and annealed to sequencing primers with reagents from an Illumina HiSeq PE Cluster Kit v4-cBot. Following transfer of the flowcell to an Illumina HiSeq instrument at the University of Utah High-Throughput Genomics Core, a 125 cycle paired-end sequence run was performed using HiSeq SBS Kit v4 sequencing reagents with six samples multiplexed in each Illumina lane. The average number of reads per sample ranged from 38 to 58 million reads.
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