Chromium™ Single Cell 3' Library & Gel Bead Kit v2, 16 rxns PN-120237 Product Components Part Number Chromium Single Cell Gene Expression provides single cell transcriptome 3' gene expression and multiomic capabilities to profile tens of thousands of cells. 4. 1000121, qty: 3; and (3) single index kit t set a, july 1, 2020: $68,483 Chromium Next GEM Chip G Single Cell Kit (1000127, 16rxns | 1000120, 48 rxns) Dual Index Kit NT Set A (1000242, 96 rxns - Protein Libraries) Dual Index Kit TT Set A (1000215, 96 rxns - Gene Expression Libraries) Targeted Gene Expression 10x Compatible BioLegend TotalSeq™-B antibodies Workflow 1 Prepare your sample CHROMIUM NEXT GEM SINGLE CELL 5' KIT V2, 16 RXNS; DUAL INDEX KIT TT SET A 96 RXNS; AND CHROMIUM NEXT GEM CHIP K SINGLE CELL KIT, 48 RXNS Solicitation ID/Procurement Identifier: RFQ-NIAID-2101571 Ultimate Completion Date: Wed Dec 21 18:00:00 GMT 2022 Chromium Single Cell Reagents With our reagent kits, capture and amplify mRNA, profile the immune repertoire, identify CRISPR perturbations, detect chromatin accessibility, and more. scRNA Seq was conducted as described previously (Becker et al. 1000286 - Chromium Next GEM Chip K Single Cell Kit, 48 rxns, Quantity 1. Cells were processed using the 10x Genomics Chromium Controller and the Chromium Next GEM Single Cell 3′ GEM, Library & Gel Bead Kit v3.1 (PN-1000121, 10x Genomics Inc.) following the manufacturer's protocol (CG000204 Rev. Single-cell capturing and library preparation were performed using the following reagents: Chromium Next GEM Single Cell 5′ Library & Gel Bead Kit v1.1, 16 rxns, PN-1000165; Chromium Next GEM Chip G Single Cell Kit, 48 rxns, PN-1000120; Chromium Single Cell V(D)J Enrichment Kit, Human T Cell, 96 rxns, PN-1000005; Single Index Kit T Set A, 96 . Read lengths: 50 x 8 x 16 x 49. For single-cell RNA sequencing, blood was obtained from patients and centrifuged on a Ficoll gradient, and mononuclear cells (whole or monocyte enriched) were used for droplet single-cell RNA sequencing (with Chromium Controller with Chromium Next GEM Single Cell 3' GEM, Library and Gel Bead Kit version 3.1 reagents [10X Genomics]), as . Our suite of Chromium Single Cell products can capture molecular readouts of cell activity in multiple . Germinal centres (GC) are lymphoid structures in which B cells acquire affinity-enhancing somatic hypermutations (SHM), with surviving clones differentiating into memory B cells (MBCs) and. The Chromium Controller uses advanced microfluidics to perform single cell partitioning and barcoding in a matter of minutes. When do I need an additional Single Cell 5' Library Construction Kit (PN-1000020) to run my experiment with Single Cell VDJ v1 or v1.1 reagent kits? Chromium Next GEM Chip K Single Cell Kit, and Library Construction Kit) enable the joining profiling of T cell receptor (TCR) specificity and gene expression at single cell resolution. Characterize perturbations at an even deeper level with Single Cell 5' Gene Expression, enabling detection of up to four modalities at single cell resolution, including guide RNAs (A), whole transcriptome gene expression (B), cell surface protein expression (C), and immune receptor sequences (D). *Note that Library Construction Kits are only sold in whole number units. Briefly, the captured cells were lysed, and the released RNA was barcoded via reverse transcription in individual GEMs. Explore cellular heterogeneity, novel targets, and biomarkers with combined gene expression, surface protein expression, or CRISPR edits in each cell. Click here for the Chromium™ Controller Firmware Update Procedure The National Institute of Allergy and Infectious Diseases (NIAID) has a requirement for: 1) Chromium Next GEM Chip K Single Cell Kit, 16 rxns CATALOG # 1000287 QTY: 1 2) Chromium Next GEM Single Cell 5' Kit v2, 16 rxns CATALOG # 1000263 QTY: 1 3) Chromium Next GEM Single Cell 3' GEM, Library & Gel Bead Kit v3.1, 16 rxns CATALOG # 1000121 QTY: 1 4) Chromium Next GEM Chip G Single Cell Kit, 16 . Click here for the Chromium™ Controller Firmware Update Procedure Chromium Next GEM Single Cell ATAC Library & Gel Bead Kit v1.1 (Cat#1000175, 10x Genomics), Chromium Next GEM Chip H Single Cell Kit (Cat#1000162, 10x Genomics) and Single Index Kit N Set A (Cat#1000212, 10x Genomics) were purchased for scATAC library preparation. Chromium Next GEM Automated Single Cell 5ʹ Kit v2, 4 rxns PN-1000298 Chromium Automated Single Cell Human TCR Amplification & Library Construction Kit, 24 rxns PN-1000300 Chromium Automated Single Cell Mouse TCR Amplification & Library Construction Kit, 24 rxns PN-1000310 Chromium™ Reagent Kits - In Brief Single Cell 3' Reagent Kit In a microfluidic Single Cell 3' Chip, a library of Single Cell 3' Gel Beads, functionalized with a 10x™ Barcoded primer, are combined with a single cell to create a single nanoliter droplet encapsulated in oil or Gel Bead-In-EMulsion (GEM). If using v2 of the Single Cell Immune Profiling Solution, proceed to the Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) with Feature Barcode technology for Cell Surface Protein & Immune Receptor Mapping user guide ( CG000330 ). Library & Gel Bead Kit v3.1, 4 rxns — 188 960,- za kus bez dph a Chromium Next GEM Chip G Single Cell Kit, 48 rxns - 47 140,- bez dph, v celkové hodnoté 285 681 KE a souhlasíme se zvetejnéním této objednávky v Registru smluv." Chromium Next GEM Chip K Single Cell Kit, 48 rxns PN-1000286 Chromium Next GEM Chip K Single Cell Kit, 16 rxns PN-1000287 Dual Index Kit TT Set A, 96 rxns PN-1000215 For version v.1.1 (Chromium Next GEM Single Cell ATAC Library and Gel Bead Kit v.1.1, 10x Genomics) prepare mastermix: 8 μl nuclei suspension (in 1× PBS + 1% BSA or 1× DNB + 2% BSA), ATAC . Generate single-cell libraries Once single-cell suspensions are isolated, they are ready for library preparation using the Chromium Next GEM Single Cell 5ʹ Kit v2. Catalog No. All samples and reagents were prepared and loaded into the chip and ran in the Chromium Controller for GEM generation and barcoding. 1000190 - Library Construction Kit, 16 rxns, Quantity 1. B, 10x Genomics Inc.). Kit Selection Guide for Chromium Connect Single cell gene expression workflows library *cDNA may be generated from Chromium X/iX, Controller or Connect, using low-throughput and standard assays for single cell gene expression. Briefly, a small volume (< 4 µl) of single-cell suspension at a density of some 2000 . Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) with Feature Barcode technology for CRISPR Screening -User Guide -Official 10x Genomics Support Chromium Next GEM Single Cell 5' Reagent Kits v2 (Dual Index) with Feature Barcode technology for CRISPR Screening User Guide, Last Modified on April 8, 2022, Permalink Request Pricing See How It Works 2. Catalog No. Library construction and single-cell RNA-seq were performed according to the instructions of the Chromium Next GEM Single Cell 3 Reagent Kit v3.1. Sequencing reads were processed by Cell Ranger version 3.0.1 (10x Genomics, Pleasanton, CA, USA) using the mouse reference transcriptome mm10. To boost yields of the CRISPR screening library, the amplified complementary DNA was amplified for an additional 21 . If you wish to make additional libraries per gel bead reaction (for . The single-cell suspension was filtered (Falcon 352235), spun at 300 g for 5 min, and resuspended in DMEM with 10% FBS. 10x Genomics Chromium Single-Cell RNA Sequencing Following FACS, cell concentrations and viability were determined on the Cell Countess II and cell suspensions (Lin−/Lin+) were loaded into the Next GEM Chip G and Chromium Controller (10x Genomics, Pleasanton, CA, USA), per manufacturer's protocol standardized for GEM formation and barcoding. Analysis and Visualization Software Convert your data to biologically meaningful insights and interactively explore your results with easy-to-use software. Products: Single Cell Immune Profiling Single-cell RNA sequencing was performed at Genomics Centre in King's College London. Recommended Sequencing Depth for Cell Surface Protein Library: Chromium Next GEM Single Cell 5'GEM Kit v2, 4 rxns PN-1000266 (store at −20°C) Chromium Next GEM Single Cell 5 ʹ GEM Kit v2 RT Reagent B 1 2000165 RT Enzyme C 1 2000102 Reducing Agent B 1 2000087 Poly-dT RT Primer 1 2000007 Cleanup Buffer 1 2000088 Amp Mix 1 2000103 cDNA Primers 1 2000089 10xGenomics.com # PN Chromium Next GEM Chip J Single Cell Kit, 16 rxns PN-1000230 Single Index Kit N Set A, 96 rxns PN-1000212 Dual Index Kit TT Set A, 96 rxns PN-1000215 Upgrading to Firmware 4.00 or higher is required before running this assay. When do I need an additional Library Construction Kit (PN-1000190) to run my experiment with Next GEM 5' v2 reagent kits? For single ATAC sequencing, libraries were generated using the 10X Chromium Next GEM Single Cell ATAC kit v1.1; Chemistry with NextGEM Chip H. The sequencing was performed on the NovaSeq 6000 using the SP flowcell 100 cycle kit. 1000215 Dual Index Kit TT Set A 96 rxns, Quantity 1. 1000120, qty: 2; (2) chromium next gem single cell 3 gem library gel be, part no. The samples were further processed using the Chromium Next GEM Single-Cell 3′ Reagent Kit (10x Genomics) according to the manufacturer's protocol (CG000184 Chromium Single Cell3 v3 Feature Barcoding CRISPR Screening UG RevB). covid-19/coronavirus - chromium cell library and kits: (1) chromium next gem chip g single cell kit 48 rxns, part no. Chromium Controller Our compact instrument Chromium Single Cell Immune Profiling reagents With our reagent kits, explore adaptive immunity through multidimensional approaches, including 5' mRNA, full-length paired BCR and TCR sequences, antigen-receptor specificity, cell surface proteins, and more. Contract Type: Lysis was diluted . Q: When do I need an additional Single Cell 5' Library Construction Kit (PN-1000020) to run my experiment with Single Cell VDJ v1 or v1.1 reagent kits? Explore cellular heterogeneity, novel targets, and biomarkers with combined gene expression, surface protein expression, or CRISPR edits in each cell. Bioinformatics Analysis of Single Cell Sequencing Data (A) Uniform manifold approximation and projection (UMAP) of scRNA-seq from bulk plasmablast with recovered BCR sequences (purple) or unrecovered (gray). 2.6. Core. A: Each Chromium Single Cell VDJ v1 or v1.1 kit contains enough library preparation materials to make one library per gel bead reaction (either gene expression, TCR or BCR). kit user guide. PN-1000128, 10X Genomics, Pleasanton, CA, USA) and Chromium Next GEM Chip G Single Cell Kit (catalog no. 10x genomics kits were used to produce the gel bead-in emulsions (gems) and the resulting sequence libraries (gene expression library, feature barcode library, tcr library) according to manufacturer's protocol: for batch i, chromium next gem single cell 5' library and gel bead kit v1.1 (10x genomics, pn-1000167), chromium next gem single cell v … Chromium Single Cell Gene Expression provides single cell transcriptome 3' gene expression and multiomic capabilities to profile tens of thousands of cells. PN-1000127, 10X Genomics . Chromium Next GEM Chip K Automated Single Cell Kit, 16 rxns 1000297 Chromium Next GEM Chip K Automated Single Cell Kit, 48 rxns 1000289 Modular workflow products Product code Chromium Next GEM Automated Single Cell 5' cDNA kit v2, 24 rxns 1000425 Automated Library Construction kit, 24 rxns 1000428 Use the filters on the left hand side to find the 10x products that match your needs. Chromium™ Single Cell 3' Reagent Kits v2 Parts from Chromium™ Single Cell 3' Reagents Kits v2 are NOT interchangable with parts from other Chromium Reagent Kits, despite the same or similar names, unless they also share the same Part Number. (C) UMAP overlay of BCR sequences with confirmed spike binding activity. Moreover, there are multiomic approaches that permit researchers at the single cell/nucleus level to quantify transcripts, chromatin accessi- bility,and,likelysoon,methylationandproteomesinalmost any sample including human biopsies (2). We describe here a protocol for rapid isolation of myeloid cells from tumor-harboring mouse cerebellum without cell sorting to minimize cell damage for scRNA-seq. 1000255 - Chromium Single Cell Mouse BCR Amplification Kit, 16 rxns, Quantity 1. During cell partitioning, each cell is combined with a single gel bead that is loaded with reverse transcription primers that include one of the 10X Barcode sequences. Chromium Next GEM Chip G Single Cell Kit, 48 rxns PN-1000120 Chromium Next GEM Chip G Single Cell Kit, 16 rxns PN-1000127 Single Index Kit T Set A, 96 rxns PN-1000213 Upgrading to Firmware 4.00 is required before running this assay. Chromium Next GEM Chip K Single Cell Kit, 48 rxns PN-1000286 (store at ambient temperature) Chromium Next GEM Chip K Single Cell Kit, 16 rxns PN-1000287 (store at ambient temperature) 10xGenomics.com Chromium Next GEM Chip K & Gaskets Chromium Next GEM Chip K 6 2000182 Gasket, 6-pack 1 370017 # PN Single cells were processed through the Chromium Single-cell Platform by using a Chromium Next GEM Single Cell 3′ GEM, Library, and Gel Bead Kit (version 3.1; catalog no. The single-cell RNA-seq (scRNA-seq) library was prepared using theChromiumSingleCell3'LibraryandGelBeadkitV2(PN-120237),Chromium Single Cell 3' Chip kit V2 (PN-120236), and Chromium i7 Multiplex kit (PN-120262)withthe10×GenomicsChromiuminstrument.Samplesweresequencedon Product Selector. NC1873088 $1,831.65 / Each Qty Add to cart This protocol includes the procedures for further enrichment of myeloid cells using CD11b+ magnetic beads, followed by the generation of scRNA library and sequencing analysis. These Single Cell RNA Sequencing of CNS Tissue. chromium next gem single cell 5' kit v2, 16 rxns; dual index kit tt set a 96 rxns; and chromium next gem chip k single cell kit, 48 rxns due: 21 dec, 2022 (in 9 months) agency: national institutes of health Single-cell RNA-sequencing analysis. SMARTer Mouse TCRa/b Profiling Kit: TaKaRa: Cat# 634402: Chromium Next GEM Chip K Single Cell Kit: 10× Genomics: Cat# 1000287: Chromium Next GEM Single Cell 5' Kit: 10× Genomics: Cat# 1000265: Chromium Single Cell Human TCR Amplification Kit: 10× Genomics: Cat# 1000252: Dual Index Kit TT set A: 10× Genomics: Cat# 1000215: Library . & gt ; 90 % viability ) were loaded onto a microfluidic Chip and ran in Chromium. X 8 x 16 x 49 Chip K Single cell chromium next gem chip k single cell kit ( catalog.. Construction and single-cell RNA-seq were performed according to the instructions of the Chromium Controller for GEM generation and.! 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