Smart seq v4 ultra low input rna kit

Manufactured by Standard BioTools

The SMART-Seq v4 Ultra Low Input RNA Kit is a laboratory equipment product designed for ultra-low input RNA sequencing. It enables the generation of high-quality cDNA libraries from as little as 10 picograms of total RNA.

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Lab products found in correlation

Seqamp dna polymerase, by Takara Bio (1 mentions) Accumax, by Innovative Cell Technologies (1 mentions) C1 single cell auto prep system, by Standard BioTools (1 mentions) Agencourt ampure xp, by Beckman Coulter (1 mentions) Nextera xt kit, by Illumina (1 mentions) Glomax, by Promega (1 mentions)

2 protocols using smart seq v4 ultra low input rna kit

Single-Cell RNA Sequencing Protocol

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LC2ad were dissociated to single cells by Accumax treatment (Innovative Cell Technologies). We employed the C1 single-cell auto prep system (Fluidigm) and SMART-Seq v4 Ultra Low Input RNA Kit with oligo DNA, identically to Smart-Seq2. We executed the C1 protocol ‘SMART-Seq v4 Rev B’. We acquired approximately 1 ng of FL-cDNA. As a control, we also conducted cDNA synthesis from bulk cells using a general thermal cycler with the same reaction conditions. Using SeqAmp DNA Polymerase (TAKARA Bio), we conducted a further amplification (1 cycle of 1 min at 96 °C, 5 cycles of 30 s at 95 °C, 65 °C for 30 s, 7 min at 68 °C, 1 cycle of 10 min at 72 °C). We purified the re-amplified cDNA using Agencourt AMPureXP (Beckman Coulter). Approximately 300 ng of FL-cDNA was obtained.

Seki M., Katsumata E., Suzuki A., Sereewattanawoot S., Sakamoto Y., Mizushima-Sugano J., Sugano S., Kohno T., Frith M.C., Tsuchihara K, & Suzuki Y. (2018). Evaluation and application of RNA-Seq by MinION. DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes, 26(1), 55-65.

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Single-Cell RNA-Seq of Sox10+ Cells

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We isolated cells from five biological samples (2x naïve and 3x H. poly infected animals (7 dpi)) from both male and female Sox10|tdT adult mice at 24 hours after tamoxifen induction. TM samples and isolation of tdT⁺ cells using FACS were prepared as described above. The quantity and quality of a single cell suspension was assessed using the Eve Cell Counter and trypan blue. 200-1000 cells were loaded into a Fluidigm C1 Single Cell Auto Prep IFC for mRNA-seq for use with the Fluidigm C1 system. After the cell load program cells were imagined using the JuliStage to identify wells that contain 1 single cell. The SMART-Seq v4 Ultra Low Input RNA Kit for the Fluidigm C1 System, IFCs was used to reverse transcribe and amplify cDNA. cDNA was quantified using the Glomax (Promega). Libraries were prepared from cDNA generated from single cells using the Nextera XT kit from Illumina. Final libraries were pooled and sequenced on the HiSeq 4000.

Progatzky F., Shapiro M., Chng S., Garcia-Cassani B., Classon C.H., Sevgi S., Laddach A., Bon-Frauches A.C., Lasrado R., Rahim M., Amaniti E.M., Boeing S., Shah K., Entwistle L.J., Suárez-Bonnet A., Wilson M.S., Stockinger B, & Pachnis V. (2021). Regulation of intestinal immunity and tissue repair by enteric glia. Nature, 599(7883), 125-130.

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