Typical applications
Whole plasmid sequencing using third generation NGS Generation technology is a powerful and accurate method for characterizing and analyzing plasmids, which make it well suited for a wide range of applications.
- Plasmid Verification
- Fast check and comparison of plasmid clones
- Resequencing of whole plasmids without a reference
How does it work?
Novel Nanopore technology is used to obtain very long sequences of several kilobases. It is a powerful method for characterising and analysing plasmids, which are small, circular pieces of DNA that can replicate independently of the genome.
There are several key benefits to using ONT for whole plasmid sequencing:
- High throughput: ONT allows for high-throughput sequencing of multiple samples in a single run, making it ideal for large-scale studies and projects.
- Long read lengths: ONT generates long, continuous reads, which are particularly useful for sequencing large plasmids or for assembling complex plasmid structures.
- High accuracy: ONT has a high accuracy rate, making it suitable for a wide range of applications, including plasmid engineering, quality control, and the identification of genetic mutations.
- High resolution: ONT provides high-resolution data, allowing researchers to accurately identify and characterize even subtle differences in plasmid sequences.
- Versatility: ONT is suitable for a wide range of sample types, including low copy number plasmids, making it a versatile tool for plasmid analysis.
Use the following concentration and volume for your samples:
|
Sample type
|
Size Category
|
Length
|
Concentration |
Min. Volume |
TAT* |
Plasmid
|
Regular |
2.5 - 25 kbp |
30 ng/µl |
20 µl |
1-2 days |
| Large |
25 - 125 kbp |
50 ng/µl |
30 µl |
5-7 days |
| XL |
125 - 300 kbp |
50 ng/µl |
50 µl |
5-7 days |
Please send your sample tubes according to the requirements below:
- Use 1.5 ml safe-lock tubes for your samples
- Please provide your samples either in nuclease-free water or elution buffer (10 mM Tris, pH 8.5)
- Please do not use Nanodrop for your concentration measurement. Spectrophotometric techniques are unreliable for the quantification measurement. Please use a fluormetric method, like Qubit.
- Do not tape or wrap tubes with parafilm. Safe-lock tubes offer perfect sealing and evaporation protection
- Label your sample tubes with our free barcode labels. Affix the label horizontally
How and where to ship your samples:
- Use either one of your nearest DropBox for free sample pick up or a courier of your choice
- You can convinently use one of our sample shipment options to send your samples
- To send your samples to us safely, make use of our sample bags and boxes
- Send your samples to our Eurofins Genomics sequencing lab in Cologne, Germany
Please ensure that your sample is free of any contaminants (e.g., ethanol) that could negatively affect your results. A complete list of contaminants can be found here: https://community.nanoporetech.com/contaminants
The following files will be delivered for each plasmid sample
- Data analysis report in HTML format
- Assembled plasmid sequence with quality scores for each nucleotide position (FASTQ)
- Assembled plasmid sequence in FASTA format
- Annotated genome sequence of all contigs in GENBANK format (import directly into your favourite software like Sanger's Artemis, SnapGene, Geneious Prime, Benchling, SeqBuilder Pro etc.)
- Per base quality with confidence score
- Raw plasmid sequence reads that aligned to the assembled plasmid as FASTQ files
- Annotated feature table, including the nucleotide sequence of each feature in TABLE format
No delivery of: Pod5 or FASTQ files that don't align.
If you are interested in Pod5 files and un-aligned FASTQ files please contact us, regarding our premium service.
What is a WPS Premium Service ?
Your samples will be run on a fresh flow cell dedicated exclusively to your samples (no pooling). After the run, the flow cell will be discarded.
When would I need to order this service ?
As described in our FAQ about index hopping, it might happen during a run that:
- A few single reads from your samples spill over into another sample
- A few single reads from other samples spill over into your samples
To completely avoid this, you should order our Premium Service. With this service, we will use one flow cell exclusively for your samples and discard it directly after use.
How can I order the Premium Service ?
The Premium Service is only available for samples submitted in plates. During order placement, please check the "Premium Service" checkbox on the order page. Please note that this service incurs an additional fee.
