Single-cell fixed RNA gene expression analysis (10× Flex)
- Overview
- Sample Submission Guidelines
- Price list
- Data Delivery
- Estimated delivery date
- Related services
Overview
This service provides single-cell fixed RNA gene expression analysis. Users submit fixed cells or nuclei samples. SignAC processes the cells or nuclei, prepares single-cell RNA-seq libraries using 10X Genomics Chromium X and Flex kits, conducts sequencing, and returns FASTQ data.
Only single plex is supported.
We use the following kits from 10x Genomics for library preparation.
・Chromium Fixed RNA Kit, Human Transcriptome, 4 rxns x 1 BC [PN-1000474] or
Chromium Fixed RNA Kit, Mouse Transcriptome, 4 rxns x 1 BC [PN-1000495]
・Chromium Next GEM Chip Q Single Cell Kit, 16 rxns [PN-1000422]
・Dual Index Kit TS Set A, 96 rxns [PN-1000251]
※ Please prepare the Chromium Next GEM Single Cell Fixed RNA Sample Preparation Kit, 16 rxns [PN-1000414] by yourself.
Sample submission guidelines
Cell pellets (prepared with the following method)
- Species: human or mouse
※Please suspend cells in quenching buffer, add 10% enhancer, further add 50% glycerol to a final concentration of 10% glycerol, and store at −80°C. (Quenching buffer and enhancer are included in the Single-Cell Fixed RNA Sample Preparation Kit.)
| Sample Type | Recommended # |
|---|---|
| Fixed cells | 200,000-2 x 106 |
| Fixed nuclei | 400,000-2 x 106 |
| FFPE dissociated suspension | 400,000-2 x 106 |
※Fixation, dissociation, and cryopreservation: Please use the 10x Genomics Chromium Next GEM Single-Cell Fixed RNA Sample Preparation kit (No. 1000414). Refer to the following links for protocols.
- Please submit an Excel file with the sample name (information written on the tube) and cell count through the Redmine system.
- Add a serial number to the sample name and write the number to the tube as well.
- Please write your name and the Redmine ticket number on your sample tube.
※Please submit your samples in 1.5 mL tubes. We recommend using DNA low-bind tubes.
Data delivery
FASTQ files
- Sequencing is targeted at >200 million reads per sample, aiming at 300 million reads.
- Please order an additional sequencing service provided by SignAC to further increase the sequencing depth after the primary data delivery.
- Please consult us for a simple primary analysis (mapping, etc.).
- In principle, libraries prepared by SignAC will not be returned.
Estimated delivery date
Please contact us.
Reference
6 weeks