Note
If submitting whole plasmid samples with Sanger sequencing samples, separate the samples in different bags and clearly mark each bag.
Sample Preparation
Sample type | Size Category | Length | Concentration | Min volume | Price per sample |
Plasmid | Regular | 2.5 - 25 kb | 30 ng/uL | ≥10 uL | $15 |
Large | 25 - 125 kb | 50 ng/uL | ≥20 uL | $30 |
XL | 125 - 300 kb | 50 ng/uL | ≥40 uL | $60 |
FAQ
- How fast is your turnaround time?
- 1 business day from when the sample arrives. Large volume orders may take a little longer.
- Is there a minimum number of samples I must submit?
- Any number of samples is welcome. There are no minimums or limitations.
- How can I ship my samples to you?
- Dropboxes are available in many locations. Also, Eurofins is the only sequencing company that offers free digital shipping labels on >95% of orders. Take a look at our sample submission page for details.
- How accurate is it?
- The equipment vendor reports accuracy of 99.3%. The research community has reported accuracy between 93-98%.
- How does whole plasmid sequencing compare to Sanger sequencing?
-
There are pros and cons of every sequencing method. Sanger sequencing and Oxford Nanopore sequencing (ONT) are both methods for determining the sequence of nucleotides in a piece of DNA. Typically Sanger is consider the most accurate method for short-read sequencing and NGS is better for long-read sequencing.
Overall, the choice between Sanger sequencing and ONT will depend on the specific needs of the application. Both methods have their strengths and limitations, and the appropriate method will depend on factors such as the length and quality of the DNA sample, the desired level of accuracy, and the cost and availability of the necessary equipment.
| Pros | Cons |
Short read sequencing (Sanger) | - Widely used and well-established method
- High accuracy and precision
- Can be automated for high-throughput sequencing
- Chromatogram makes it easy to visually interpret results
| - Limited read length (typically up to 1000 base pairs)
- Requires relatively large amounts of high-quality DNA
- Requires reassembly for covering longer regions.
- Difficult to sequence repetitive regions without gaps and resolve large variations.
|
Long read sequencing (Whole Plasmid) | - Long read lengths (up to several hundred thousand base pairs)
- Can sequence a wide range of sample types, including low-quality and degraded DNA
- No primer design required
- All data from one sample
| - Lower accuracy compared to Sanger sequencing, particularly for shorter reads
- More expensive per base pair compared to Sanger sequencing
- ·Cannot resolve single bases.
|
- How should I prepare my samples?
-
Sample type | Size Category | Length | Concentration | Min volume | Price per sample |
Plasmid | Regular | 2.5 - 25 kb | 30 ng/uL | ≥10 uL | $15 |
Large | 25 - 125 kb | 50 ng/uL | ≥20 uL | $30 |
XL | 125 - 300 kb | 50 ng/uL | ≥40 uL | $60 |
- Can I use my free barcodes on whole plasmid sequencing orders?
- Absolutely. Using our free barcode labels is not required, but always an option.
- Can I use my EVOcard to pay for my order?
- Absolutely, yes.
- Can you sequence my mixture of plasmids?
- You can send it and we can sequence it, but we cannot predict or promise the analysis outcome. The customer would take on the risk of these orders. service for plasmids is intended for a clonal population of molecules. You can send mixtures of molecular species, but since we can't predict the analysis outcome, it's at your own risk. If your species are very similar (e.g. differ by only a few nucleotides), the pipeline will most likely create a single .fasta consensus file with low confidence positions at SNP/indel locations. You can view those locations in your provided stats.csv and .fastq files. If your species are sufficiently distinct (e.g. vastly different in size or sequence), the pipeline will first attempt to make a .fasta consensus file from the highest abundance species. It will also attempt to make a consensus of other species with read counts that are >20% of that of the most abundant species. Ultimately, which species end up producing a consensus will vary depending on overall sample quality, coverage, and relative abundance/degradation of each species. Sequencing is considered successful if the pipeline is able to generate any consensus, even if it is not your target. Re-sequencing mixtures won't change the relative proportions of the species, but you can submit multiple aliquots if you need higher total coverage. If the pipeline does not produce a consensus for your target, you can download the raw reads from your dashboard and bin them yourself, but please note that raw reads are much more noisy and error-prone (~98.3% accurate) than consensus reads. If you are interested in sequencing a known mixture (e.g. barcode or variant libraries), please refer to the section below on.
- Can I order through my institutions marketplace or B2B punchout?
- We will be rolling out the new order page across portal sites, marketplaces, and punchouts very soon. At the moment, please submit orders on the main www.eurofinsgenomics.com website.
- What is the coverage?
- It really depends on the sample quality. We do not guarantee any specific level of coverage at this timeA good sample submitted at the required concentration typically yields hundreds or thousands of raw sequencing reads. Consensus coverage depends on how many reads are full-length plasmids and how many, if any, degraded. We do not guarantee any specific level of coverage at this time.
- What data files is delivered?
- Every order recieves the following files: 1.) .fasta, 2.) .gbk GeneBank file, 3.) Histogram file, 4.) .html pLannotate map. Additionally, the .fastq files can be provided on request.
- Can I submit bacterial genome samples or Amplicons?
- We will be adding this service soon but not yet.