Initial Meeting: Participate in a 30-minute meeting to present the kit and address any questions. Kit Shipment: Confirm participation, send a purchase order (PO), and receive the kit. Shipping typically takes less than a week to the USA or Europe. Kit Storage and Preparation: Store the kit properly until use. Optionally, request a protocol walk-through meeting to discuss specific applications and procedures. Wet Lab and Sequencing: Perform the wet lab and sequencing process. Send quality control (QC) of the sample and sequencing data. Bioinformatics Report: Receive the bioinformatics report within two weeks. Participate in a one-hour debriefing session to discuss results. Feedback: Complete a feedback form at the end of the program.

The single-cell kit contains two boxes with all necessary reagents for single-cell isolation, barcoding, and cDNA generation. The kit includes: Box 1: Cell partitioning chip, magnet, barcoded beads, and wash buffers A and B (stored at 2°C to 8°C). Box 2: Lysis buffer, RNase inhibitor, DTT, RT master mix, reverse transcriptase mix, TS primer, amplification enzyme, amplification master mix, and A primer mix (stored at -25°C to -15°C). Additional Services: Sequencing data processing and a bioinformatics report. Available in presentations for 2 or 16 reactions. Note: Sequencing library preparations for ONT/PacBio platforms must be ordered separately.

Specific Target: The magnetic beads specifically target poly-A tailed mRNA by having a poly-T sequence at the 3’-OH end. Number of UMI (unique molecular identifiers) per Bead: Each bead contains 10^7-10^8 unique binding sites, each associated with a unique molecular identifier (UMI). This means each bead can bind to 10^7-10^8 different mRNA molecules, each tagged with a unique UMI.

Per Chip: Each chip can partition between 3,000 to 10,000 cells by loading 15,000 to 35,000 cells. Combined Analysis: Up to 20,000 cells can be analyzed together by pooling two reactions, thanks to different barcodes on the generated cDNA.

Standard Chip Compatibility: The standard chip is compatible with cells or nucleus preparations ranging from 5 to 25 μm in size. Customized Chip Requirements: Larger cells (up to 70 μm) or irregularly shaped cells require a customized chip. Sample Preparation Recommendations: Fresh cell samples with high viability in PBS solution are recommended. Security Features: The chip is disposable, making it suitable for handling high-security samples, such as those used in HIV investigations. Successfully Tested Cell Types: Examples of compatible cell types include PBMC, ZebraFish embryo cells, HEK-293, NIH-3T3, MDA-MB-453, K-562, A549, and PC3, among others.

Yes, the chip can be operated manually without any special equipment. You only need a regular P200 pipette to load the chip.

The width of the tip is crucial for chip loading, and using the right tip can significantly facilitate loading. Here are some pipette tips that have been validated for successful chip loading: Eppendorf cat #0030078551 Jet Biofil PMT231200 VWR cat #732-3701 Vertex #F42L7N0F-VXRG Rainin cat#30389242 Extragene TS250CFS Opentrons OT-2 #999-00081 For first-time users, it is recommended to use some of these P200 tips. If you have a different brand of pipettes or need further guidance on tip selection, you can consult with us for advice on the best tip to use for your specific setup.

Visual Monitoring: The microwell chip is designed as an open system, allowing users to visually monitor the entire process. Magnification: The process can be observed with a 10X magnification, providing clear visibility of cell and bead loading. Transparency and Control: The design enables step-by-step observation, offering transparency and control over each stage of the workflow.

Consumables: Regular sterile RNAse/DNAse-free certified filter tips 1.5 mL nucleic acid low-binding tubes PCR tubes Equipment: Single channel pipettes Cell counter or inverted microscope and hemocytometer Mini-centrifuge Vortex Thermal mixer Thermal cycler DynaMag™ magnetic rack QuBit™ 4 fluorometer or equivalent Nucleic acid fragment analyzer ONT/PacBio Sequencer Reagents: Absolute ethanol Nuclease-free water RNase AWAY™ or equivalent 10 mM Tris-HCl pH 8.0 10% Tween-20 PBS pH 7.4 AMPure XP or PB QuBit™ dsDNA quantitative kit or equivalent Library prep and sequencing reagents

General Compatibility: The kit is specifically designed for long-read sequencing platforms and is compatible with both PacBio and ONT technologies. This allows for the use of the same cDNA preparation for both platforms. Library Preparation Recommendations: For PacBio, Kinnex Full Length RNA is recommended. For ONT, LSK 114 is recommended. ONT Single-Cell Experiments: For single-cell experiments using ONT, it is recommended to use a P2 or superior equipment. Data should be basecalled in SUP or HAC mode for optimal results.

We will return your data in a format compatible with common downstream bioinformatics tools (such as FLAMES or IsoSeq), along with a bioinformatics report, within two weeks. The bioinformatics report includes: Sequencing and sample statistics Pipeline information Details on computational resources used Overview of read structure (to check for chimeric reads or other artifcats) Cell calling metrics Seurat plots SQANTI3 plots Please contact us if you are interested in customized data processing.

Data Submission: You will need to provide raw sequencing data for demultiplexing using ArgenTag's proprietary barcode calling software. Confidentiality: Your data will be kept confidential and not shared or used for unauthorized purposes, as outlined in our terms and conditions. If you are still unable or unwilling to share your sequencing data (e.g., for human samples), please stay tuned: a standalone, user-installable version of our barcode calling software is currently under development and is expected to be released in 2025.