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.

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.

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.

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.

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.

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.

Technology Platforms

Clonal Sequencing

• Oxford Nanopore MinION
• Novasesq 6000
• Illumina Nextseq500
• Illumina Miseq

Sample QC

• Qubit Fluorometer
• Agilent 2100 Bioanalyzer
• Agilent 2200 TapeStation
• Agilent Fragment Analyzer Systems

QPCR

• Applied Biosystems Quantstudio 3
• Applied Biosystems Quantstudio 5
• Fluidigm Biomark HD

Droplet Digital PCR

• BioRad QX200 

Single Cell Prep Systems

• 10X Genomics Chromium 

High Throughput Compound Handling

• PHERAstar FSX HTS Microplate Reader
• Eppendorf epMotion 5070
• Agilent Bravo
• Formulatrix Mantis
• Labcyte Access/Echo550

Sonication

• Diagenode Bioruptor Pico