Molecular Biology 

The Molecular Biology Core provides a wide range of services for precision and high throughput analysis of DNA and RNA to assist in molecular biology projects within the CRUK Manchester Institute. We have developed a multitude of workflows for automated assay running and support scientists with  state-of-the-art analytical instrumentation.

image of a lab

 

Clonal Sequencing Service

This comprehensive service covers all aspects of Next Generation Sequencing (NGS). An extensive portfolio of library prep methods for RNA and DNA sequencing is set up to cope with a wide range of input quantity and quality. The bulk of sample input is processed by automated workflows using high throughput instrumentation, e.g. Agilent Bravo, Formulatrix Mantis. The majority of the sequencing is performed on Illumina NGS instruments, including Miseq and Novaseq 6000. In addition, we have validated a number of applications on Oxford Nanopore long read sequencing platforms.

Agilent Bravo automation platform for NGS library prep

 

Illumina Novaseq 6000 clonal sequencing platform

 

Novaseq 6000 S4 flow cell

 

Oxford Nanopore Long Read Sequencing

 

Cell Line Authentication and Pathogen Screening

To maintain the highest standards of experimental work the CRUK Manchester Institute encourages regular testing of cell lines for their authenticity and has a strict policy on Mycoplasma control. For correctly identifying cell lines MBC provides a NGS based Cell Line Authentication Assay which detects highly variable short tandem repeat (STR) regions in the genome. Test results are cross-checked against an extensive in-house cell line database as well as ATCC references.

Prior to injection into mice all cell line samples must be screened for mycoplasma and Murine Hepatitis Virus (MHV). MBC provides a regular PCR based screening service to facilitate compliance.

MiSeq FGx platform for STR profiling

 

qPCR and digital PCR Instrumentation

The Service supports researchers with a set of 96-well and 384-well instruments (Thermofisher Quantstudio) for quantitative real-time PCR and houses a high precision robot (Eppendorf Epmotion) for fast reagent dispensing. We also support an integrated fluid circuit (IFC) real-time PCR platform (Fluidigm Biomark HD) for high throughput qPCR. In addition, a droplet digital PCR system (Bio-Rad QX200) is installed for absolute quantitation of DNA or RNA.

QS3 and QS5 qPCR instruments

 

Biorad QX200 digital PCR platform

 

Single Cell and Spatial Transcriptomics

Using the 10X Genomics Chromium platform for single cell encapsulation and processing, MBC supports methodologies for single cell genomics, transcriptomics, and multiseq applications.

A recently emerging and rapidly developing field, we adopted applications for spatial transcriptomics analysis as well as multiomics data integration.

10X Genomics Chromium platform for single cell encapsulation and processing

 

Visium technology for spatial transcriptomics analysis

 

High Throughput Compound Handling and Screening Service

Automated compound handling is performed on a Echo550 (Beckman Coulter) acoustic dispenser. This is incorporated into the Access robotic platform that further integrates a thermocycler (Inheco) as well as a microplate reader (BMG Pherastar FSX) for high throughput plate assaying. The setup is used for customized, high precision compound dispensing as well as for single and combinatorial drug dosing experiments. In addition, bespoke sets of compound libraries, including kinase inhibitor as well as FDA approved drug libraries, are  available for cell screening purposes.

The Labcyte Access platform incorporates the Echo550 Acoustic compound dispenser and microplate reader.

 

Technology Platforms


Clonal Sequencing

Sample QC

QPCR

Droplet Digital PCR

Single Cell Prep Systems

High Throughput Compound Handling

Sonication