|Sample type||Category||Size||Concentration||Minimum volume||Price per sample||Order now|
|Linear/Amplicon||Standard||600 bp - 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|
The linear/amplicon sequencing service is intended for the sequencing of clonal linear DNA from 600 bp to 125 kb in length. This service is performed using the newest long-read sequencing technology from Oxford Nanopore Technologies (ONT), and includes the following components:
You will receive the following data for whole plasmid sequencing:
We also return two files that show how confident we are in our basecall at each position of our consensus sequence:
This service requires 300 ng (standard linear/amplicon) or 1000 ng (big linear/amplicon) of linear double-stranded DNA, normalized to the specific concentration listed in the table above.
Submit the final purified linear/amplicons 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.
Linear/amplicon 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 or big linear/amplicon 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!
This service is intended for linear double-stranded DNA molecules.
Size verification should be performed on full-length linear/amplicon DNA via gel electrophoresis.
The linear/amplicon workflow is more tolerant to degradation than the plasmid workflow because it does include some minimal fragmentation during the library prep process (and thus we do not return read length histograms for linear/amplicon samples because a clonal peak is not produced and it is typically not informative). However, for best results, you'll still want to aim for intact linear DNA molecules.
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 molecule. Sending mixtures of molecular species will give mixed results and is at your own risk!
Please refer to our shipping instructions for details.