Sequencing and Identification
At NCIMB we have extensive experience of identifying unknown bacterial and fungal isolates. We can search sequences for genes associated with particular products and properties, rapidly quantify organisms within samples and undertake microbial community analysis.
We provide a microbial identification service and partner with companies, universities and research organisations to carry out research and development projects.
- Fast, confidential and accurate identification to GMP standards.
- A range of turnaround times.
- Clear, easy to understand reporting, tailored to your requirements.
- Easy sample submission.
We undertake rapid genotypic identification of bacterial isolates, including mycoplasma, by sequencing the 16S ribosomal DNA gene with full gene or 500bp options available.
Sequenced data is examined against the validated MicroSEQ® database. When this doesn’t provide a high enough species level match we can use the more comprehensive, non-validated, European Nucleotide Archive (ENA) database from EMBL-EBI, the NCIMB reference collection and the client’s own historic data collection to match sequences to published data.
As custodians of the UK’s National Collection of Industrial, Food and Marine Bacteria, we have immediate access to this unique source of authentic reference material. We also have access to a number of private collections, ensuring a comprehensive coverage of genera and species.
Reports can be tailored to your requirements and include top sequence matches and phylogenetic trees.
Strain to strain comparison and differentiation
Comparison of isolates at the strain level can be valuable with respect to the patented use of bacteria and production of many food products.
We can use two different approaches:
- 16S rDNA sequencing can be used for basic strain comparison when the full 16s (1500bp) is sequenced. An alignment of several sequences highlights major differences, but does not always distinguish between closely related strains.
- Multilocus sequence typing (MLST) is a highly reproducible DNA sequencing-based technique. Sequences of internal fragments of (typically) seven essential house-keeping genes, i.e. those essential for cells to function, are used to characterise isolates.
- MLST via NGS - this approach uses whole genome sequencing data to perform MLST on a genome wide scale.
Fungal identification – yeasts and moulds
There are two distinct approaches to genotypic identification of fungi: sequencing of the D2 region of the large subunit ribosomal DNA (D2 LSU), and sequencing of either one or both of the internal transcribed spacer regions between the small and large subunit ribosomal RNA genes (ITS). At NCIMB we combine these techniques to offer a comprehensive service that gives our customers the most reliable species level identification available.
For D2 LSU identification, sequences are analysed against the validated MicroSEQ® database. In some cases, however, this only gives a genus level identification. When this occurs we often find that if it is required a species level identification can be obtained by using the ITS method.
Quantitative polymerase chain reaction (qPCR)
NCIMB offers a qPCR service. Quantitative polymerase chain reaction is a valuable means of determining the presence or absence of an organism or group of organisms in a sample, whilst also providing very rapid quantitative information.
We can develop customised qPCR assays for specific organisms of interest to individual clients or undertake assays for a range of commonly requested organisms.
High throughput sequencing
Microbial community analysis
NCIMB can undertake microbial community analysis. This technique, which has revolutionised understanding of environmental microbiology, uses sequencing information to analyse the whole microbial population from a single sample. It identifies the groups of microorganisms present to give a comprehensive picture of the microbial ecosystem.
Whole genome sequencing
Advances in sequencing technology have paved the way to screening bacteria to identify those that could be useful in the development of new applications and improved microbial processes. At NCIMB we can carry out data analysis of whole genome sequences to identify gene sequences associated with potentially useful products and properties.
Pure cultures can be sent to us for identification as plates, slopes or broths that have been incubated to produce sufficient growth. Please complete a sequencing request form and send with isolates to be identified.
We can also accept mixed cultures and other sample formats, such as samples of water, slimes, raw materials or contaminated products, but please contact us prior to submitting them for identification.
NCIMB does not accept active cultures of dangerous human pathogens or organisms isolated from clinical or diseased plant materials for identification. We cannot accept isolates known to be greater than ACDP category 2 and ACGM class 1.
Take care to package samples appropriately. Detailed advice on the packaging of isolates is available on request.