|Sample type||Category||Size||Concentration||Minimum volume||Price per sample||Order now|
|Plasmid||Standard||2.5 - 25 kb||30 ng/uL||≥10 uL||$15 or 1 dinocoin||Order|
|Big||25 - 125 kb||50 ng/uL||≥20 uL||$30 or 2 dinocoins||Order|
|Huge||125 - 300 kb||50 ng/uL||≥40 uL||$60 or 4 dinocoins||Order|
The whole plasmid sequencing service is intended for the full-length sequencing and annotation of clonal circular plasmid DNA between 2.5 kb and 300 kb in length. This service is performed using the newest long-read sequencing technology from Oxford Nanopore Technologies (ONT), and includes the following components:
In the vast majority of cases, we deliver plasmid sequencing results within one business day of receipt of your samples.
You will receive the following data for whole plasmid sequencing:
Finally, we return two files that show how confident we are in our basecall at each position of our consensus sequence:
Unlike traditional Sanger sequencing, which relies on primers to detect only a specific small region, our
full-length ONT service sequences each entire plasmid molecule with a single long read. All molecules
a received sample are sequenced, including any degraded plasmids or background genomic DNA; we do not use
any primers which would target specific regions or types of molecules.
As a result:
This service requires 300 ng (standard plasmid), 1000 ng (big plasmid), or 2000 ng (huge plasmid) of circular, double-stranded DNA, normalized to the specific concentration listed in the table above.
Please refer to published literature for plasmid extraction protocols. Submit the final purified plasmids in elution buffer (10 mM Tris, pH 8.5) or nuclease-free water; avoid buffers containing EDTA (e.g. TE or AE buffer) whenever possible.
Plasmid samples are sequenced WITHOUT primers, so please DO NOT ship any primers with your samples or mixed into your samples.
Our low sequencing prices and fast turnaround times do not include DNA extraction or quality control (QC) services, so please verify with full QC that your samples meet the following requirements prior to shipping.
Submit your standard, big, or huge plasmid samples normalized to the specific concentration and
volume listed in the table above. Quantification assay must be performed with Qubit
fluorometric method (such as a plate reader).
IMPORTANT: Do not use Nanodrop for DNA quantification – spectrophotometric methods are NOT reliable for quantification!
Sending samples at too high OR too low concentration may adversely affect the library prep and/or sequencing reactions, possibly resulting in sequencing failure. Sending samples at the incorrect concentration (usually due to using Nanodrop for quantification) is by far the most common cause of sequencing failure. Accurate quantification and normalization are key!
For best results, aim for intact circular double-stranded plasmids. Plasmids that are degraded or fragmented are much more likely to result in sequencing failure by yielding no consensus due to lack of full-length sequencing reads.
Size verification should be performed on full-length plasmids (NOT digested or amplified plasmids) via gel electrophoresis; use a linear ladder for linearized plasmids, and a supercoiled ladder for intact circular plasmids.
We recommend samples with 260/280 above 1.8 and 260/230 between 2.0-2.2. Purity assay may be
or other spectrophotometric methods.
IMPORTANT: Do not refer to the DNA concentration reported by Nanodrop as a substitute for the above fluorometric quantification!
For best results, samples should NOT contain any of the following:
Samples should also be "pure" in the sense that they should only contain copies of a single clonal plasmid molecule. Sending mixtures of molecular species will give mixed results and is at your own risk!
Please refer to our shipping instructions for details.
Our Dropboxes and the free & reduced cost UPS labels generated during order submission both provide overnight shipping within the US to our Louisville lab location, and both provide next-morning results for whole plasmid and linear/amplicon sequencing. We recommend that US customers use either a Dropbox or a plasmidsaurus-generated UPS label whenever possible (rather than self-shipping).
International Dropbox shipments are usually delivered to us within 2-3 days and we provide results on the same morning of receipt. If you do not have a local dropbox, you can self-ship via any carrier to our Louisville lab location:
4743 Poplar Level Road
Louisville, KY 40213
Phone #: 541-514-0871
Please ensure any required forms are included with your shipment.
NOTE ABOUT DROPBOXES: The specific day that each dropbox ships out varies by site, and may not be every weekday for your local dropbox. Please check the dropbox list to find the shipping day(s) for your local dropbox! To ship your samples on a different day, please refer to "Alternative Shipping Options" in the detailed shipping table. Contact us at email@example.com if you’re interested in hosting a new dropbox at your location!