Sre0036

Manufactured by Merck Group

Sourced in United States

SRE0036 is a laboratory equipment product from Merck Group. It is designed for general laboratory applications. The core function of SRE0036 is to provide a reliable and consistent platform for various laboratory procedures. No further details are available without the risk of bias or extrapolation.

Automatically generated - may contain errors

Lab products found in correlation

Nextseq 500, by Illumina (2 mentions) Chromium controller, by 10x Genomics (1 mentions) Red blood cell lysis buffer, by Abcam (1 mentions) Tissuelyser lt, by Qiagen (1 mentions) Novaseq 6000 sequencer, by Illumina (1 mentions)

5 protocols using sre0036

Single Cell RNA-seq of TdTomato+ Cells

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FACS sorted TdTomato+ cells were counted and diluted in 1X PBS with 0.04% BSA (Sigma Cat# SRE0036) before being loaded onto the 10X Genomics Chromium instrument. The scRNA-seq libraries were generated using the 10X Chromium Single Cell 3′ v2 reagent kit according to the manufacturer’s instructions and were sequenced on an Illumina Nextseq500.

Rueda E.M., Hall B.M., Hill M.C., Swinton P.G., Tong X., Martin J.F, & Poché R.A. (2019). The Hippo Pathway Blocks Mammalian Retinal Müller Glial Cell Reprogramming. Cell reports, 27(6), 1637-1649.e6.

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Dissociation and Single-Cell RNA Sequencing

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Samples were finely minced with a scalpel and digested with 0.25% Trypsin-EDTA (ThermoFisher #25200056) for 30 minutes at 37°C with occasional agitation. The digestion was terminated by additional of RPMI (ThermoFisher #R8758) supplemented with 10% Fetal Bovine Serum (ThermoFisher # 16000044) and the cell suspension was filtered with a 70 μm mesh. A cell pellet was collected following 5 minutes of centrifugation at 300 ⨯ g. Red blood cells were removed with Red Blood Cell lysis buffer (BioVision, #5830-100). The cells were resuspended in PBS + 0.04% BSA (Sigma-Aldrich, #SRE0036), counted and 3,000 − 5,000 cells were loaded into the chromium controller (10X Genomics) following the manufacturer's instructions and processed using version 3 of the 3' scRNA-seq protocol (Suppl. Table S1).

Nowicki-Osuch K., Zhuang L., Cheung T.S., Black E.L., Masqué-Soler N., Devonshire G., Redmond A.M., Freeman A., di Pietro M., Pilonis N., Januszewicz W., O'Donovan M., Tavaré S., Shields J.D, & Fitzgerald R.C. (2023). Single-Cell RNA Sequencing Unifies Developmental Programs of Esophageal and Gastric Intestinal Metaplasia. Cancer Discovery, 13(6), 1346-1363.

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Single Cell RNA-seq of TdTomato+ Cells

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Quantifying Influenza Lung Viral Titers

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To assess lung viral titers, lung tissue was collected in 1 mL of DPBS and subject to homogenization in Qiagen Tissue LyserLT (Qiagen, United States) using Lysing Matric C tubes with beads (116912050-CF; MPBiomedicals, United States). Plaque assays was performed on MCDK cells with MEM overlay containing 1% oxoid agar, 0.125% BSA (SRE0036; Sigma, United States) and 2 μg N acetylated trypsin (T6763; Sigma, United States). Plaques were visualized by crystal violet. The limit of detection of the influenza plaque assays was 50 pfu/lung tissue or 50 pfu/mL cell supernatant.

Anand G., Clark-Dinovo C., Perry A.M., Goodwin V.M., St. Raymond E., Sakleshpur S, & Steed A.L. (2024). Aromatic amino acid metabolites alter interferon signaling and influenza pathogenesis. Frontiers in Molecular Biosciences, 10, 1232573.

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Single-Cell RNA-Seq of 3D Spheroids

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3D spheroids were cultured for 9 days. Single-cell suspensions were obtained by washing once with PBS and mechanically dissociating with TrypLE Express. Cell suspensions for each cell line were combined to obtain a cell suspension of 1x10 6 cells/mL in PBS + 0.04% BSA (Sigma #SRE0036). Cell suspensions were processed for single-cell RNA-seq libraries using the 10X Genomics Chromium platform according to the 3' RNAseq V2 library kit instructions. Sequencing was performed on an Illumina NovaSeq 6000 sequencer according to standard protocols.
Sequenced scRNA-seq libraries were processed using the 10x Cell Ranger Pipeline 2.0.1 by mapping to the hg19 genome. The average mRNA expression level from each hallmark gene set in MSigDB cell was analyzed using the Single-Cell Signature Explorer 85 to calculate an overall single-cell expression intensity score for each gene set.

Čermáková K., Smith E.A., Chan Y.S., Cabrera M.L., Chambers C., Jarvis M., Simon L.M., Xu Y., Jain A.K., Putluri N., Chen R., Ripley R.T., Veiseh O., & Hodges H.C. (2021). SMARCA4 regulates spatially restricted metabolic plasticity in 3D multicellular tissue.

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