Massive Parallel Sequencing
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Massive Parallel Sequencing (MPS), also known as Next Generation Sequencing (NGS) is the collection of technologies that are able to genotype (determine the sequence) of DNA or RNA fragments in a massive and parallel fashion.
The second generation MPS technologies rely on clonal amplification of the DNA/RNA fragments to be genotyped, whilst third generation MPS technologies can directly sequence single DNA/RNA fragments without prior clonal amplification.
The second generation technologies generate short sequences of the clonally amplified fragments and are referred to as short-read sequencers. They rely on sequencing-by-synthesis (SBS), where the stepwise incorporation of nucleotides - starting from a sequencing primer - is detected through changes in either fluorescence, pH or impedance. These technologies generally have a low error rate.
The third generation technologies generate long sequences and are referred to as long-read sequencers. The sequence of the DNA (or RNA) fragment to be inferred, is established by measuring either (1) the potential difference over a membrane when this fragment migrates through an nanopore embedded in this membrane or (2) the real-time incorporation of fluorescently labelled nucleic acids by a polymerase. Generally, these technologies have a higher error rate than the 2nd generation sequencing technologies.
Specifications & applications
Flow cell types
R9.4.1
R10.4
Standard v2
Standard v3
Nano v2
Micro v2
SP
S1
S2
S4
Max. read length
> 4 Mb
250 bp
300 bp
250 bp
150 bp
250 bp
150 bp
150 bp
150 bp
Max. run time
72h (interruptible at all time)
39h
56h
28h
19h
38h
25h
36h
44h
Max. capacity (bases)
< 50 Gb
8,5 Gb
15 Gb
0,5 Gb
1,2 Gb
400 Gb
500 Gb
1250 Gb
3000 Gb
Max. capacity (reads)*
< 5M (for avg. 10K bp reads)
15M SE - 30M PE
25M SE - 50M PE
1M SE - 2M PE
4M SE - 8M PE
0,8B SE - 1,6B PE
1,6B SE - 3,2B PE
4,1B SE - 8,2B PE
10B SE - 20B PE
* Abbreviations used : SR = Single Read ; PE = Paired End ; M = million ; B = Billion ; Gb = Gigabases
** To know more about the Q score metric, see below
Sample types
PCR products (amplicons) > DNA
Microtube* or 96 well plate
See test details/specifications
Room temperature
Blood > DNA
EDTA tube
10 ml (min. 3 ml)
Room temperature
Blood > RNA
PAXgene Blood RNA tube
10 ml (2,5 ml blood)
Ice pack
Blood > cfDNA/ctDNA
Streck Cell-Free DNA BCT
9 ml
Room temperature
Blood > cfRNA/exoRNA
Streck RNA Complete BCT
10 ml
Room temperature
Cells of various tissue origins > DNA
Microtube* with cell pellet or on lysis buffer
> 5 million cells
Dry ice
Cells of various tissue origins > RNA
Microtube* with cells on lysis buffer
> 5 million cells
Dry ice
DNA from various tissues > DNA
Microtube* or 96 well plate
See test details/specifications
Room temperature
Blood > DNA
DBS / Guthrie card / Punch in microtube*
Min. 1 punch
Room temperature
FFPE block of various tissues > DNA
Paraffin block (in cassette)
Representative block ; Tumor load preferentially >10% ; To be processed by our pathology department (microtome section needed)
Room temperature
FFPE block of various tissues > RNA
Paraffin block (in cassette)
Representative block ; Tumor load preferentially >10% ; To be processed by our pathology department (microtome section needed)
Room temperature
FFPE section of various tissues > DNA
Microtube*
Min. 1x 10 µm section (up to 2 mm³ tissue) ; Tumor load preferentially >10%
Room temperature
FFPE section of various tissues > RNA
Microtube*
Min. 1x 10 µm section (up to 2 mm³ tissue) ; Tumor load preferentially >10%
Room temperature
NGS library (non-PCR free) > DNA
Microtube* or 96 well plate
See test details/specifications
Room temperature or ice pack
NGS library (PCR free) > DNA
Microtube* or 96 well plate
See test details/specifications
Ice (max. 1 day) or dry ice
Plasma > cfDNA/ctDNA
DNA LoBind 5 ml tube
> 2ml
Dry ice
Plasma > cfRNA/exoRNA
DNA LoBind 5 ml tube
> 2ml
Dry ice
RNA from various tissues > RNA
Microtube*
See test details/specifications
Ice (max. 20 min) or dry ice
RNA from various tissues > RNA
Microtube* or 96 well plate
See test details/specifications
Ice (max. 20 min) or dry ice
Sars-Cov2 RNA (naso/oropharyngeal) > RNA
Microtube* or 96 wel plate (plate is preferred)
Log(copies/ml) > 3 (min. 10 µl)
Ice (max. 20 min) or dry ice
Extra requirements
See test/application specific recommendations
:
More information can be found on the subpages (related tests)
Deliverables
Below, you can find the type of data files that can be retrieved for this test.
If you require more information or need a more custom output, please consult our Bioinformatics page.
Run folder
The complete run folder can be transferred if it concerns a run reservation (private run)
RUO (on request)
Raw data - fast5 file
Raw data file generated by Oxford NanoPore sequencers
RUO (on request)
Raw data - bam file
Binary data file containing the aligned reads
RUO (on request)
Documents & certificates
141-MED (UZ Brussel)
245-MED (Hopital Erasme)
Sciensano recognized
141-TEST (BRIGHTcore)
EFI
Plan experiment
There are 2 options available. See below for more details.
Run reservation
Have full control and produce your own libraries. We will sequence them for you.
To start a reservation you need to get in touch with us to (1) clarify your needs (2) get a specific quote and (3) deliver your libraries.
1
Application
Choose a specific application from our portfolio and let us process your samples
Check our applications below. You will find more information on the application pages. Get in touch with us if you don't find what you need.
2
Related tests
Through gene panel sequencing, a subset of genes can be selected for genotyping. This reduces cost and as a result unlocks the ability for deep-sequencing.
For germline genetic testing, full human genome sequencing is becoming mainstream. Find out more here.
Identify fetal aneuploidies and even microdeletions and -duplications in the maternal blood
Perform transcriptome studies on a wide variety of input materials.
Sequence the full Sars-Cov2 genome to assess the evolution of the virus within the population
Identify single cell aneuploidies, or gene expression or the combination of both.
Only interested in coding genes and not willing to bear the cost of WGS? An exome might be your assay!
Identify SNVs, CNVs and even SVs by sequencing the entire genome