Sample Type | Category | Size | Concentration | Minimum Volume | Price Per Sample (USD) |
---|---|---|---|---|---|
Bacterial Genome | Standard | < 7 Mb | 50 ng/μL | 20 μl | $90 |
Big | 7 - 12 Mb | 50 ng/μL | 20 μl | $105 | |
Standard with Extraction | < 7 Mb | See Instructions Below | $105 | ||
Big with Extraction | 7 - 12 Mb | See Instructions Below | $120 | ||
Hybrid | < 7 Mb | 50 ng/μL | 20 μl | $165 | |
Big Hybrid | 7 - 12 Mb | 50 ng/μL | 20 μl | $195 | |
Hybrid with Extraction | < 7 Mb | See Instructions Below | $180 | ||
Big Hybrid with Extraction | 7 - 12 Mb | See Instructions Below | $210 |
The bacterial genome sequencing service is intended for whole-genome sequencing and assembly of genomic DNA (gDNA) from a clonal population (single species) of bacteria.
When you send pre-extracted gDNA, we deliver bacterial genome sequencing results within 1-2 business days of receipt of your samples. When our genomic DNA extraction option is selected, we deliver bacterial genome sequencing results within 3-5 business days of receipt of your samples. When the Hybrid sequencing option is selected, we deliver hybrid results with 6-8 business days (or 8-10 business days if you also include the extraction option).
This service is performed using the latest long-read sequencing technology from Oxford Nanopore Technologies (ONT), and includes the following components:
When your results are ready, you will receive an email notification. Once you sign in to your account, you can download these results from your Dashboard. More detailed information about data & file types can be found in the Results Interpretation Guide.
The target output of this service is a high-accuracy, full-length contig for each chromosome and plasmid for your bacterial species, with a set of bacterial genome annotations. However, our ability to deliver the target output is directly dependent on the quantity, quality, and purity of the gDNA or bacterial cells sent to us. We do not guarantee any specific output.
Successful sequencing is defined by achieving at least one of the following deliverables:
If we are not able to achieve at least one of those deliverables, then our repeat policy applies. Even when a high-quality assembly cannot be generated, we still provide the raw data and the report, and you may also still receive some of the other file types.
We are now offering a Hybrid ONT + Illumina bacterial genome sequencing service that polishes your long-read ONT assembly with Illumina reads. We use the Polypolish v0.6 tool to improve the consensus quality at those tricky homopolymer runs and methylation motifs. The options for this service (standard and big genomes, with and without extraction) are listed in the order table on the Dashboard.
In addition to the ONT-only target deliverables listed above (ONT-only assembly and files), this hybrid service adds on these additional target deliverables:
The input DNA requirements for this hybrid service are identical to the ONT-only requirements, as detailed below. You do not need to send extra DNA!
This service requires 1 µg of high quality, high purity, high molecular weight (HMW), double-stranded gDNA, with recommended >50% of the DNA above 15 kb in length and recommended purity of 260/280 above 1.8 and 260/230 between 2.0-2.2. Our low sequencing prices and fast turnaround times do not include gDNA quality control (QC) services, so it is your responsibility to verify that you are preparing suitable samples that meet these requirements prior to shipping them to us.
There are numerous approaches to bacterial culture, thus we do not provide specific recommendations. Please refer to published literature for protocols.
Any extraction method that yields high quality, high purity, high molecular weight (HMW), double-stranded gDNA that is free of nicks, gaps, breaks, and contaminants is suitable for this sequencing service.
Our in-house preferred methods for extraction are the Zymo® Quick-DNA Miniprep Plus Kit (with a lysozyme pre-treatment for gram positive samples) and Zymo® Quick-DNA Fungal/Bacteria Miniprep Kit. Here are a few other extraction methods that we and others have had success with:
How to handle gDNA samples:
All samples must be sent as cell pellets that have been resuspended in Zymo 1X DNA/RNA Shield preservative. Please do not use any other type of preservation media, such as RNAlater, as they do not preserve the DNA as effectively and do not fully inactivate the host cells, which is required for safe handling in our lab. We accept cell pellets from both BSL1 and BSL2 strains.
We strongly recommend growing a fresh clonal culture of your bacteria in liquid broth and harvesting the cells when they are in exponential growth or early stationary phase. Please do not send cells from older cultures that are in the death phase.
NOTE: Our extraction service does not include growing your samples; we extract DNA directly from the material you send us. It is important that enough cells have been collected, otherwise the DNA extraction will likely fail.
Due to customs regulations, the bacterial extraction service is currently only available to US and UK customers.
1. Pellet the cells by centrifugation and remove as much
supernatant as possible.
We require the equivalent of 8-12
OD600 or 4-6 x 109 cells (e.g. 8-12 mL culture at
1.0 OD600).
Please place an order, either for Standard with extraction (up to 7 Mb) or Big with extraction (7-12 Mb).
When filling out your sample names, please add either a plus (+) or a minus (-) at the end of each sample name to indicate whether each strain is gram positive or gram negative. Not doing so will cause a processing delay for your order, as we need this information directly from you.
Please label each 2 mL screw cap tube with the
order code, sample number, and the plus (+) or a minus (-).
For example, "X2X1-" would indicate order code X2X, sample 1, gram
negative.
To prevent damage and leakage of the tubes during shipment, please place the screw cap tubes in a rigid container such as a falcon tube or tube box before shipping. For orders of more than 10 samples, please put the tubes in a tube box and load the samples row by row in numerical order as this greatly reduces the handling time when receiving your samples.
Please send preserved cell pellets at room temperature. Dry ice and cold packs are not necessary.
Please refer to our Results Interpretation Guide for details.