What sequencing platform is used?

For all of our sequencing we are using the latest Oxford nanopore chemistry, achieving >99% single-read accuracy (the “Q20 chemistry”) and Q50 consensus accuracy.

More information: https://nanoporetech.com/platform/accuracy

What data is provided?

Along with the assembled fasta sequence of your sample, we provide read length distribution histogram, assembly coverage graph, sequence annotation file and variant call table.

What are the general recommendations for DNA samples?

We highly recommend DNA samples to be purified to a high purity, for example silica column based purification. Running agarose gel is optimal to ensure quality. Fluorimetry based DNA qualifications methods (for example the Qubit fluorimeter) are superior to absorbance based (for example nanodrop).

Can the sequencing fail?

In rare cases the sequencing is unsuccessful. There is no single reason for this to happen, it is usually a combination of smaller unfortunes, including lower DNA concentration, genomic DNA contamination, suboptimal storage conditions, DNA fragmentation. We are multiplexing samples and they are intrinsically prone to slightly different outputs.

More questions?

How many reads can I expect per sample for a standard plasmid or amplicon run?

In most cases, we generate from 100 to a few thousand reads per sample. Typically, 20-30 full-length reads are sufficient to return a high-quality sequence of a standard plasmid sample.

What if I need more reads?

If you require more reads, a simple solution is to prepare and submit additional tubes containing the same DNA sample and request the fastq files. This allows us to pool the data during analysis to increase coverage.

Can you sequence non-standard plasmid DNA?

Yes, with some considerations:

• For plasmids 30-100 kb in size: if your plasmid sample is larger than standard synthetic plasmids (>30kb), contains impurities (for example was extracted from a native host, is a single-copy plasmid), or is contaminated with genomic DNA, we recommend submitting multiple tubes of the same reaction. Please indicate in the submission form that reads from multiple samples should be pooled before the analysis.

• For plasmids larger than 100 kb or complex mixtures: we recommend using our small genome sequencing service for optimal results.