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200 cycle kit

Manufactured by Illumina
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The 200 cycle kit is a consumable product designed for use with Illumina's sequencing platforms. It provides the necessary reagents and materials to perform up to 200 cycles of sequencing on a single sample. The kit's core function is to enable the sequencing process, facilitating the determination of the nucleotide sequence in a DNA or RNA sample.

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Lab products found in correlation

Nextera xt kit, by Illumina (3 mentions) Nextseq, by Illumina (3 mentions) Mantis, by Formulatrix (2 mentions) Tpplabtech mosquito hts, by Formulatrix (2 mentions) 2100 bioanalyzer, by Agilent Technologies (2 mentions) Novaseq, by Illumina (1 mentions) Mosquito hts, by Formulatrix (1 mentions) Rnaeasy micro kit, by Qiagen (1 mentions) Ovation solo rna seq system, by Tecan (1 mentions) Novaseq 6000, by Illumina (1 mentions) Allprep dna rna mini kit, by Qiagen (1 mentions) Truseq stranded total rna kit, by Illumina (1 mentions) Novaseq 6000 apparatus, by Illumina (1 mentions)

Spelling variants of this product

TruSeq® SBS v3–HS 200 cycle kit (8 citations) TruSeq 200 cycle SBS kit (6 citations) HiSeq Rapid SBS Kit v2 200 cycles (4 citations) NovaSeq 6000 S1 Reagent Kit - 200 cycle (3 citations) TruSeq Rapid SBS Kits–200 Cycle (2 citations) Novaseq S4 200 cycles v1.5 kit (2 citations) TruSeq SBS Kit v3 - HS (200-cycles) reagents (2 citations) TruSeq SBSHS Kit v3 200 cycles (2 citations) NovaSeq 6000 S4 Reagent Kit (200 Cycles) (2 citations)

5 protocols using 200 cycle kit

1

Single-Cell RNA-seq Using Smart-seq-2

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Cell lysis, and cDNA synthesis was performed using the Smart-seq-2 protocol as described previously 68 ,69 , with some modifications. After cDNA amplification (23 cycles), cDNA concentrations were determined via capillary electrophoresis and cells were cherry-picked to improve quality and cost of sequencing. Cell selection was done through custom scripts and simultaneously normalizes cDNA concentrations to ~0.2 ng/uL per sample, using the TPPLabtech Mosquito HTS and Mantis (Formulatrix) robotic platforms. Libraries were prepared using the Illumina Nextera XT kits following the manufacturer’s instructions. Libraries were then sequenced on the Nextseq (Illumina) using 2 × 75bp paired-end reads and 2 × 8bp index reads with a 200 cycle kit (Illumina, 20012861) and pooled using the Mosquito liquid handler. Library quality was assessed via capillary electrophoresis on a Fragment Analyzer (AATI) and quantified by qPCR. Samples were sequenced at an average of 700,000 reads per cell.
Yousef H., Czupalla C.J., Lee D., Chen M.B., Burke A.N., Zera K.A., Zandstra J., Berber E., Lehallier B., Mathur V., Nair R.V., Bonanno L.N., Yang A.C., Peterson T., Hadeiba H., Merkel T., Körbelin J., Schwaninger M., Buckwalter M.S., Quake S.R., Butcher E.C, & Wyss-Coray T. (2019). Aged blood impairs hippocampal neural precursor activity and activates microglia via brain endothelial cell VCAM1. Nature medicine, 25(6), 988-1000.
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2

Smart-seq-2 Modified Single-cell RNA-seq

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Cell lysis, first-strand synthesis and cDNA synthesis was performed using the Smart-seq-2 protocol as described previously (Picelli et al., 2014 (link)) in both 96-well and 384-well formats, with some modifications. After cDNA amplification (22 cycles), cDNA concentrations were determined via capillary electrophoresis (96-well format) or the PicoGreen quantitation assay (384-well format) and wells were cherry-picked to improve quality and cost of sequencing. Cell selection was done through custom scripts and simultaneously normalizes cDNA concentrations to ~0.2 ng/μL per sample, using the TPPLabtech Mosquito HTS and Mantis (Formulatrix) robotic platforms. Libraries were prepared and pooled using the Illumina Nextera XT kits and in-house Tn5, following the manufacturer’s instructions. Libraries were then sequenced on the Nextseq or Novaseq (Illumina) platforms using 2 × 75bp paired-end reads and 2 × 8bp index reads with a 200-cycle kit (Illumina, 20012861). Samples were sequenced at an average of 1.5M reads per cell.
Chen M.B., Yang A.C., Yousef H., Lee D., Chen W., Schaum N., Lehallier B., Quake S.R, & Wyss-Coray T. (2020). Brain Endothelial Cells Are Exquisite Sensors of Age-Related Circulatory Cues. Cell reports, 30(13), 4418-4432.e4.
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3

Single-cell RNA-seq profiling of T-cell subsets

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RNA derived from sorted cell populations (Vγ9Vδ2, nonVγ9Vδ2 γδ, αβ) was isolated using the RNAeasy micro kit (Qiagen, Cat. No./ID: 74004). RNA quality was checked using a Bioanalyzer 2100 (Agilent Technologies). Indexed cDNA libraries were obtained using the Ovation Solo RNA-Seq System (NuGen) following the manufacturer’s recommendation. The multiplexed libraries were loaded on a NovaSeq 6000 (Illumina) using an S2 flow cell, and sequences were produced using a 200 Cycle Kit (Illumina, PN: 20028313). Paired-end reads were mapped against the human reference genome GRCh38 using STAR software (version 2.7.10a) to generate read alignments for each sample. Annotations Homo_sapiens.GRCh38.90.gtf were obtained from ftp. Ensembl.org. After transcript assembling, gene level counts were obtained using HTSeqd software. Differential expression was performed by using EdgeR quasi-likelihood running under the Degust platform. Only genes with a minimum count per million of 1 in each replicate were included. Volcano plots were generated using EnhancedVolcano R package (v1.10).
Sanchez Sanchez G., Papadopoulou M., Azouz A., Tafesse Y., Mishra A., Chan J.K., Fan Y., Verdebout I., Porco S., Libert F., Ginhoux F., Vandekerckhove B., Goriely S, & Vermijlen D. (2022). Identification of distinct functional thymic programming of fetal and pediatric human γδ thymocytes via single-cell analysis. Nature Communications, 13, 5842.
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4

Single-Cell RNA-seq Using Smart-seq-2

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Cell lysis, and cDNA synthesis was performed using the Smart-seq-2 protocol as described previously 68 ,69 , with some modifications. After cDNA amplification (23 cycles), cDNA concentrations were determined via capillary electrophoresis and cells were cherry-picked to improve quality and cost of sequencing. Cell selection was done through custom scripts and simultaneously normalizes cDNA concentrations to ~0.2 ng/uL per sample, using the TPPLabtech Mosquito HTS and Mantis (Formulatrix) robotic platforms. Libraries were prepared using the Illumina Nextera XT kits following the manufacturer’s instructions. Libraries were then sequenced on the Nextseq (Illumina) using 2 × 75bp paired-end reads and 2 × 8bp index reads with a 200 cycle kit (Illumina, 20012861) and pooled using the Mosquito liquid handler. Library quality was assessed via capillary electrophoresis on a Fragment Analyzer (AATI) and quantified by qPCR. Samples were sequenced at an average of 700,000 reads per cell.
Yousef H., Czupalla C.J., Lee D., Chen M.B., Burke A.N., Zera K.A., Zandstra J., Berber E., Lehallier B., Mathur V., Nair R.V., Bonanno L.N., Yang A.C., Peterson T., Hadeiba H., Merkel T., Körbelin J., Schwaninger M., Buckwalter M.S., Quake S.R., Butcher E.C, & Wyss-Coray T. (2019). Aged blood impairs hippocampal neural precursor activity and activates microglia via brain endothelial cell VCAM1. Nature medicine, 25(6), 988-1000.
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5

RNA Extraction and Sequencing from Tumor and Normal Tissues

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RNA was extracted from frozen tumor and normal tissue using the AllPrep DNA/RNA Mini kit (Qiagen, Germany) according to the manufacturer’s instructions. RNA quality was assessed using a Bioanalyzer 2100 (Agilent Technologies). A total of 22 patients had sufficient tumor RNA quantity from both pre- and posttreatment timepoints. A total of 11 patients had sufficient RNA quantity in normal tissue samples from both pre- and posttreatment timepoints. Among the patients without enough RNA quantity in normal tissue, six had biopsies containing mainly fatty tissue without any epithelial cell. Indexed cDNA libraries were obtained using the TruSeq Stranded Total RNA Kit (Illumina) following the manufacturer’s recommendations. The multiplexed libraries were loaded onto a NovaSeq 6000 apparatus (Illumina) using a S2 flow cell and sequences were produced using a 200 Cycle Kit (Illumina).
Gómez-Aleza C., Nguyen B., Yoldi G., Ciscar M., Barranco A., Hernández-Jiménez E., Maetens M., Salgado R., Zafeiroglou M., Pellegrini P., Venet D., Garaud S., Trinidad E.M., Benítez S., Vuylsteke P., Polastro L., Wildiers H., Simon P., Lindeman G., Larsimont D., Van den Eynden G., Velghe C., Rothé F., Willard-Gallo K., Michiels S., Muñoz P., Walzer T., Planelles L., Penninger J., Azim HA J.r., Loi S., Piccart M., Sotiriou C, & González-Suárez E. (2020). Inhibition of RANK signaling in breast cancer induces an anti-tumor immune response orchestrated by CD8+ T cells. Nature Communications, 11, 6335.
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