Long-read sequencing technologies

Long-read sequencing technologies

PacBio and Oxford Nanopore sequencing

Long-read sequencing can determine the nucleotide sequence of long sequences of DNA, roughly between tens and hundreds of thousands of base pairs. This removes the need to cut and amplify DNA (which is a standard procedure for short-read sequencing by Illumina).

There are several long-read sequencing technologies commerically available and continuously being developed. Of these, we offer the following two:

 

Technology PacBio Oxford Nanopore (ONT)
Applications Genome assembly - Analysis of repeats, structural variations - Sequencing of longer amplicons - Full-length transcripts, alternative splicing Genome assembly
Project type Can be combined with short reads by Illumina. Rather larger projects with a higher price. We highly recommend to combine ONT with short-reads by Illumina. Rather small to medium projects with a lower total cost.
Available sequencing settings Variable, see our pricelist or contact us
Capacity Sequel II: Highly variable (50-150 Gb very approx.)* MinION: Highly variable (1-10 Gb approx.)
Sequencing errors Low error rate in HiFi mode, higher error rate in CLR mode. Random errors.
Relatively high-error rate. Systematic errors.
Guarantee The amount of data output is approximate.
Library preparation We do not allow preparation of libraries by our clients.
Library type No limits
Outputs Standard (commonly used data formats etc.)

 

* PacBio technology is capable of producing up to several hundred kilobases of sequence length, and sequencing accuracy can be as high as 99.5%. Unfortunately, the maximum length and quality cannot be achieved at the same time. If the user seeks high-quality data, approx. 15kb long reads can be obtained. On the other hand, achieving several tens to hundreds of kilobases long reads is possible, but with a reading accuracy of only approx. 90-95%.




Case study - genome assembly

In our lab these technologies are primarily requested by customers interested in assembling new genomes (but not necessarily).

Genome assembly is the computational process of deciphering the sequence composition of DNA within the cell of an organism, using short or long (or both) sequencing reads. It can be described as solving a jigsaw puzzle. The DNA fragments (pieces) produced by long-read sequencing are easier to assemble into a complete DNA sequence simply because they are longer, same as jigsaw puzzle with less larger pieces.

This makes the long-read sequencing the technology of choice for this task. However, due to lower data quality of long-read technologies overall it is beneficial to think of combining them with Illumina short reads. Read about genome assembly by using data from Oxford Nanopore and Illumina technologies!


Libraries

For PacBio/ONT technologies, we do not allow preparation of libraries by our clients. You must supply DNA/RNA.


Results, guarantee and data analysis

Once the sequencing run is finished, you will receive its standard outputs from us. Because each sequencing project has specific data analysis requirements, we offer turnkey data analysis for the needs of your project - Custom Data Analysis. Our bioinformaticians can help you with any data analysis goal. If you are interested in this service, individual consultation is required, we recommend you seek our help already at the project preparation stage.


Processing time

Approx. 6 weeks (data analysis not included)


Course or workshop

If you are new to NGS, we encourage you to attend our two-day NGS introductory course. We will be happy to welcome you on some of other courses and workshops too.


Sample requirements

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