School of Biomedical Sciences
生物醫學學院
The Chinese University of Hong Kong 香港中文大學

Single Cell Omics Core

About us

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The SCOC provides a streamlined, cell-to-sequencing library service package for Single Cell RNA sequencing (scRNA-seq). It is capable to simultaneously profiling of transcriptomes ranging 500 – 10,000 cells in a single experiment. Different indexing systems are designed to label cell, transcript and sample origin in the sequencing library. This enables multiplexing of different sequencing libraries from multiple samples in the same Next Generation Sequencing (NGS) run. Libraries are Illumina-format compatible.

The service provided by SCOC includes an initial sample QC check, following by an evaluation of overall quality of sample for fitness to continue run. Single cells are then encapsulated in thousands of Gel Bead-in Emulsion (GEM) droplets, where cells are lysed and mRNA molecules reverse transcribed into cDNA. cDNA with specific cell barcodes are then pooled together and subjected to subsequent conventional sequencing library preparation. A final QC is performed on the final sequencing library to ensure sound quality for massively-parallel sequencing.

The SCOC welcomes interested PIs for experimental troubleshooting, intellectual advices as well as project discussions / collaborations.

Equipment

SCOC equipment
  • Countess 3 Cell Counter (Cell viability check and counting)
  • Light Microscope / Hemocytometer (Manuel cell counting)
  • Compact cooling centrifuge
  • 10X Controller (Single cell encapsulation)
  • PCR workstation
  • PCRs, thermomixer, vortex, plate centrifuge
  • Tapestation 4200 (cDNA & sequencing library QC)
  • Qubit 4 Fluorometer (DNA library quantification)

Service Workflow

Workflow overview

Workflow Overview

10X Chromium Next GEM Single Cell 3’ Gene Expression Workflow

Next GEM Workflow

Chromium Next GEM Chip:
Next GEM Chip

  • Up to 8 samples per chip processed in parallel
  • 500 to 10,000 cells per channel
  • 18 minutes run time per chip
  • Preferred cell size < 30 µm
  • ~60-65 % cell recovery efficiency

RT, template switching
RT, template switching

Pre-amp PCR
Pre-amp PCR

cDNA quality control
cDNA quality control

Sequencing library preparation
Sequencing library preparation

Sequencing library quality control
Sequencing library Control

Service request

Please contact Dr Joaquim Vong (email: Email住址會使用灌水程式保護機制。你需要啟動Javascript才能觀看它 / 39430318) for service request, technical enquiry and project discussion.

For service request, please fill in our Sample submission form.

Service fees include: (please enquire)

✓ Package set: From cell to sequencing library
✓ Cell quality and viability check / basic QC
✓ Sequencing library QC and quantification
✓ Intellectual advices (wet / dry lab)
✓ Troubleshooting

× Sequencing NOT included
× Repeated initial sample QC checks might incur extra charges

Output items

A. Sequencing library
B. QC reports for cDNA and sequencing library
C. Indexing information

The final product is a double-stranded cDNA molecule with sequencing adaptors and 3 different indices (10X barcode: cell info; UMI: transcript info; sample index: sample info). The sequencing library quality is evaluated by running into an Agilent 4200 TapeStation System.

10X Barcode
Source: 10X Genomics

Precautions

Users are required to fill in and sign the “Service Form for Single Cell Omics Core” prior to sample submission
https://webapp.sbs.cuhk.edu.hk/eform/view.php?id=55400

All users MUST declare the nature of sample(s) submitted to SCOC FREE of any bio-hazardous material.

An initial assessment of cell quality will be conducted for the generation of data with optimal quality. Users may take the risk of sub-optimal data quality if the initial cell quality check fails.

Upon service booking, a copy of the confirmed service form will be forwarded to both the end user as well as the corresponding PI.

Initial sample submission evaluation (sample quality, amount, etc) is recommended to optimize sample quality, depending on the sample nature. Potential experimental discussion(s) might be arranged prior to samples submission.

The complete workflow take approximately 2 full working days. As only limited amount of samples can be performed per week, users are encouraged to optimize their sample quality before submission. Service bookings are arranged on a first-come-first-serve basis.

Once the service has started, service charge would be accounted for FULL service package price, as the reagents are on package stock basis. We are pledged to provide our service in a reasonable, efficient manner with quality.

Useful Resources

10X Genomics – Single Cell Gene Expression
https://www.10xgenomics.com/products/single-cell-gene-expression

10X University – A step-by-step learning and training environment
https://www.10xgenomics.com/10x-university

10X Datasets
https://www.10xgenomics.com/resources/datasets

10X Publications
https://www.10xgenomics.com/resources/publications

Worthington Tissue Dissociation Guide
http://www.worthington-biochem.com/tissuedissociation

Contacts

Dr Joaquim Vong  
Scientific Officer  
Email: Email住址會使用灌水程式保護機制。你需要啟動Javascript才能觀看它
Tel no: 39430318
   
Miss Kathy Sham  
Technician  
Email: Email住址會使用灌水程式保護機制。你需要啟動Javascript才能觀看它
Tel no: 39439859
   
Location: LIBSB Room 525
Lab no.: 39434373