Nextseq 500 550 mid output kit v2.5 150 cycles

Manufactured by Illumina

The NextSeq 500/550 Mid Output Kit v2.5 (150 Cycles) is a sequencing reagent kit designed for use with the Illumina NextSeq 500 and NextSeq 550 sequencing systems. The kit provides the necessary reagents to perform 150 cycles of sequencing.

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Lab products found in correlation

Qubit dsdna hs kit, by Thermo Fisher Scientific (1 mentions) Nextseq 550 instrument, by Illumina (1 mentions) Nanodrop 2000c spectrophotometer, by Thermo Fisher Scientific (1 mentions) 2100 bioanalyzer, by Agilent Technologies (1 mentions) High sensitivity dna kit, by Agilent Technologies (1 mentions) Qiazol, by Qiagen (1 mentions) Rneasy plus universal mini kit, by Qiagen (1 mentions) Truseq stranded mrna lt kit, by Illumina (1 mentions) Nextseq 550, by Illumina (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions) Immunoseq human tcrb kit, by Adaptive Biotechnologies (1 mentions)

Spelling variants of this product

NextSeq 500 (5724 citations) NextSeq 550 (932 citations) NextSeq 500/550 (98 citations) NextSeq 500 Mid Output v2 kit (16 citations) Mid Output Kit (15 citations) NextSeq 500 Mid Output kit (14 citations) NextSeq 500/550 Mid-Output v2 (150 cycle) Kit (3 citations) 150-cycle mid output kit (3 citations) Mid Output v2.5 kit (2 citations) Kit v2 (2 citations)

3 protocols using nextseq 500 550 mid output kit v2.5 150 cycles

RNA-Seq Analysis of Gene Knockdown in HEK293T Cells

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Total RNA was extracted from four biological replicates each of WT, NT, KD_2, and KD_3 HEK293T cells using QIAzol (Qiagen, catalog# 79306) and RNeasy Plus Universal Mini Kit (Qiagen, catalog# 73404) as per manufacturer’s instruction. RNA concentration was found out using a Nanodrop 2000c spectrophotometer (Thermo Fisher Scientific), and the cDNA library was synthesized from 300 ng of RNA using QuantSeq 3′ mRNA-Seq Library Prep Kit FWD for Illumina (Lexogen, catalog# 015.96) as per the manufacturer’s instruction. The concentration of the cDNA library was assessed using Qubit dsDNA HS kit (Invitrogen, catalog# Q32851) on a Qubit four Fluorimeter (Thermo Fisher Scientific). The average size of the library was assessed using a High Sensitivity DNA Kit (Agilent, catalog# 5067–4626) on a 2100 Bioanalyzer (Agilent). Equal amounts of the libraries were pooled, and 2 nmoles was sequenced using NextSeq 500/550 Mid Output Kit v2.5 (150 Cycles) (Illumina, catalog# 20024904) on NextSeq 550 instrument (Illumina). The read was single-ended and 76 bp long. The raw sequence data is deposited in the Gene Expression Omnibus (NCBI) repository (accession ID: GSE196907) and is publicly available.

Rajendran A., Soory A., Khandelwal N., Ratnaparkhi G, & Kamat S.S. (2022). A multi-omics analysis reveals that the lysine deacetylase ABHD14B influences glucose metabolism in mammals. The Journal of Biological Chemistry, 298(7), 102128.

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Transcriptome Profiling of GFP-Labeled Tumor Tissue

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Fluorescence microscopy was used to microdissect GFP-expressing tissue from age matched tumors at P65 and P95 time points (n=3 per experimental group × n=3 animals per group). RNA was isolated using the RNeasy Plus Mini Kit (Qiagen, 74134) according to the manufacturer’s protocol. RNA integrity (RIN ≥ 8.0) was confirmed using the High Sensitivity RNA Analysis Kit (AATI, DNF-472-0500) on a 12-Capillary Fragment Analyzer. Illumina sequencing libraries with 6 bp single indices were constructed from 1 μg total RNA using the TruSeq Stranded mRNA LT kit (Illumina, RS-122-2101). Resulting libraries were validated using the Standard Sensitivity NGS Fragment Analysis Kit (AATI, DNF-473-0500) on a 12-Capillary Fragment Analyzer. Equal concentrations (2nM) of libraries were pooled and sequenced with a sequencing depth of approximately 20 million reads per sample using the NextSeq 500/550 Mid Output Kit v2.5 (150 Cycles) (Illumina, 20024904) on an Illumina NextSeq550.

Curry R.N., Aiba I., Meyer J., Lozzi B., Ko Y., McDonald M.F., Rosenbaum A., Cervantes A., Huang-Hobbs E., Cocito C., Greenfield J.P., Jalali A., Gavvala J., Mohila C., Harmanci A.S., Noebels J., Rao G, & Deneen B. (2023). Glioma epileptiform activity and progression are driven by IGSF3-mediated potassium dysregulation. Neuron, 111(5), 682-695.e9.

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TCRB Repertoire Profiling by immunoSEQ

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After PicoGreen quantification and quality control by Agilent BioAnalyser, 188-200ng of genomic DNA were split equally into six reactions and prepared using the immunoSEQ human TCRB Kit (Adaptive Biotechnologies) according to the manufacturer’s instructions. Briefly, multiplex PCR was used to amplify the CDR3 region for 31 cycles. After clean-up, 2μL of PCR product was used as input into library preparation with 8 cycles of PCR. Barcoded samples were pooled by volume and sequenced using custom primers on a NextSeq 500 in a SR155 run, using the NextSeq 500/550 Mid Output Kit v2.5 (150 Cycles) (Illumina). The loading concentration was 1pM and 20% spike-in of PhiX was added to the run to increase diversity and for quality control purposes. Raw BCL files were transferred to the immunoSEQ Analyser for processing and analysis.

Jain N., Zhao Z., Feucht J., Koche R., Iyer A., Dobrin A., Mansilla-Soto J., Yang J., Zhan Y., Lopez M., Gunset G, & Sadelain M. (2023). TET2 guards against unchecked BATF3-induced CAR T cell expansion. Nature, 615(7951), 315-322.

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