|Sample type||Category||Size||Concentration||Minimum volume||Price per sample||Order now|
|Bacterial Genome||Standard||up to 7 Mb||50 ng/uL||≥20 uL||$90 or 6 dinocoins||Order|
|Big||7 - 12 Mb||50 ng/uL||≥20 uL||$105 or 7 dinocoins||Order|
|Standard with extraction||up to 7 Mb||Send a bacterial pellet in DNA/RNA Shield||$105 or 7 dinocoins||Order|
|Big with extraction||7 - 12 Mb||Send a bacterial pellet in DNA/RNA Shield||$120 or 8 dinocoins||Order|
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. This service is performed using the latest long-read sequencing technology from Oxford Nanopore Technologies (ONT), and includes the following components:
In the vast majority of cases, 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 1 week of receipt of your samples.
You will receive the following data for bacterial genome sequencing:
The target output of this service is a single high-accuracy, circular (when appropriate for the species), full-length bacterial chromosome contig of your bacterial species, with a set of bacterial genome annotations. However, our ability to deliver target output is directly dependent on the quantity, quality, and purity of the gDNA sent to us. We do not guarantee any specific output.
If we are not able to obtain:
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 method for extraction is the Zymo® Quick-DNA Miniprep Plus Kit (with a lysozyme pre-treatment for gram positive samples). 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 DNA/RNA Shield. We accept both BSL1 and BSL2 strains.
We do not grow your samples, we extract the DNA directly from the material you send us. It is important that enough cells have been collected, otherwise the DNA extraction will likely fail.
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. Do not send cells from older cultures that are in death phase.
We require the equivalent of 8-12 OD600nm or 4-6 x 10^9 cells (e.g. 8-12 mL culture at 1.0 OD600nm). A compact cell pellet with all the supernatant removed (e.g. E. coli pellets) should be approximately 15 mg. A wet cell pellet where all the supernatant cannot be removed without disturbing the pellet (e.g. Streptococcus sp. pellets) should be approximately 30-50 mg. DO NOT send more than 50 mg of material or there will be too many cells to be protected from degradation by the DNA/RNA Shield.
The cells should be pelleted by centrifugation, washed with 1 mL PBS and pelleted again.
Place an order on our website and receive a 3-character code. When filling out your sample names please be sure to indicate if your strain is gram positive or gram negative by adding either a plus (+) or a minus (-) at the end of your sample name. Failure to do so may result in failure of the DNA extraction.
Resuspend the final pellet in 0.5 mL of 1X DNA/RNA Shield in a 2 mL screw cap tube. Please label each tube with the order label and sample number and either a plus sign (+) or a minus sign (-) to indicate if the strain is gram positive or negative. For example, "X2X1-", "X2X2-", etc,
To prevent damage and leakage of the tubes during shipment, we ask that you put the tubes in a rigid container such as a falcon tube or tube box before shipping. For orders of more than 10 samples we ask that you 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 samples at room temperature.
Due to customs regulations, the bacterial extraction service is currently only available to US customers.
Please refer to our shipping instructions for details.