Sequencing only
Sequencing with Illumina technology (MiSeq or Novaseq)
High quality input samples are key to obtaining good results. Once libraries are prepared, there is very little that can be done to improve the quality of results. Therefore, please read the following instructions carefully.
Important note: If you have prepared and pooled your libraries, quantification should be checked via qPCR and Tapestation, Bioanalyser or equivalent, prior to library submission. This is to ensure that the material you submit contains correctly ligated adaptors and does not contain excess adaptor dimers. We accept no responsibility for incorrect loading due to inefficient library construction.
We utilize the Illumina MiSeq for small scale projects as well as testing pooled libraries prior to loading on the Illumina NovaSeq 6000. It is capable of automated single end or paired-end reads using a total of 50, 300, 500 or 600 cycles.
Typical applications:
- Targeted gene sequencing
- Small genomes
- Amplicon sequencing
- 16S, 18S or ITS metagenomics
MiSeq run options
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MiSeq Reagent Kit v3 |
MiSeq Reagent Kit v2 |
MiSeq Reagent Kit v2 Micro |
MiSeq Reagent Kit v2 Nano |
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2 × 75 bp |
2 × 600 bp |
2 × 25 bp |
2 × 150 bp |
2 × 330 bp |
2 × 150 bp |
2 × 250 bp |
2 × 150 bp |
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20-24 million |
10-12 million |
4 million |
1 million |
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2.8-3.3Gb |
10–13 Gb |
750–850 Mb |
4.5–5.1 Gb |
6–8 Gb |
1.2 Gb |
500 Mb |
300 Mb |
N.B. Low diversity samples such as amplicons, including 16S, 18S, ITS and CO1, require much lower loading concentration so the output and number of reads would typically be reduced to 30%-50% of those quoted above.
The Illumina NovaSeq 6000 provides a versatile platform to support a wide variety of applications from bacterial and fungal pathogenomics to whole human genome sequencing. This is achieved using multiple flow cell types and read length combinations. The SP, S1, and S2 flow cells provide quick and powerful sequencing for the majority of high-throughput applications. The NovaSeq S4 flow cell enables the highest throughput of sequencing at the most cost-effective price for a range of applications.
Typical applications:
- De novo genome sequencing (small to large genomes)
- Genome re-sequencing
- Amplicon sequencing
- RNAseq transcriptome sequencing
- Epigenetics such as ChIP-Seq, ATAC-Seq, Whole Genome Bisulphite Sequencing (WGBS)
- Metagenomics
- Single cell sequencing
Flowcell output / scalability:
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SP |
S1 |
S2 |
S4 |
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Cycle options |
650–800 M |
1.3–1.6 B |
3.3 B–4.1 B |
8-10 B |
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100 cycle SR or PE |
65-80 Gb |
134–167 Gb |
333–417 Gb |
N/A ‡ |
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200 cycles SR or PE |
134-167 Gb |
266–333 Gb |
667–833 Gb |
1600–2000 Gb |
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2 x 150 bp PE |
200–250 Gb |
400–500 Gb |
1000–1250 Gb |
2400–3000 Gb |
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2 x 250 bp PE |
325-400 Gb |
N/A |
N/A |
N/A |
Sequencing with Nanopore Technology
Oxford Nanopore Technologies, as the name suggests, use biological nanopores embedded in silicon for long read sequencing of DNA and RNA. Sequencing is performed by the nucleic acid passing through the nanopore, causing changes in current depending on the sequence of bases that go through. Realtime computation can then call the bases (and potentially epigenetic base modifications such as methylation too).
We offer sequencing on the MinION for standard scale experiments and the PromethION for larger scale sequencing. Most applications will benefit from shearing to between 5-20kb which increases the molarity and throughput. However, longer reads in the order of hundreds of kilobases or higher are possible but with reduced yield.
Scalability overview (from nanopore website):
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Flongle |
MinION Mk1B |
PromethION 24 |
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Number of channels per flow cell |
126 |
512 |
3000 |
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Maximum number of flow cells per device run (Max flowcells with live basecalling) |
1 |
1 |
4-24 |
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Run time |
1 min - 16 hours |
1 min - 48 hours |
1 min - 72 hours |
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DNA sequencing yield per flow cell (typical in field - best in field (BIF). Yields are also dependent on chosen sample and preparation methods) |
0.5 - 2 Gb |
15 - 30 Gb |
50 - 180 Gb |
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Suitable applications include |
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