Frequently asked questions

Need a quote to generate a PO or to share with colleagues?

For our standard services and orders with fewer than 24 samples, you can generate a formal quote directly through our Price & Order page. 

To generate a quote, simply select your requested services and click “Add to Cart”. After verifying your service selection, click “Quote or Checkout”.

Here, you will have the option to either create a quote or continue to checkout via PO or credit card.

The quote will be loaded directly into your SeqCoast account, where you have the option to email the quote to yourself or others.

For custom services or bulk volume orders, please contact us for a quote!

Please see our Sample Preparation Guide (below) for detailed information about how your samples should be prepared.

  • Please see our Shipping page for detailed information about how to ship your samples to us.

When items are added to your cart, a sample manifest is automatically generated with your order information. If possible, please complete and upload this sample manifest at the time of order submission.

Alternatively, download a blank copy of our Sample Manifest Template and email the completed sample manifest to scientists [at] seqcoast [dot] com.

  • DNA extraction: Let us do the hard work for you! We offer DNA extraction using bead beating lysis. If you want to tackle it yourself, many column-based extraction kits provide good results, as do other methods if care is taken to remove contaminants. Our library prep methods work best with buffers containing less than 1mM EDTA.
  • RNA extraction: we recommend Qiagen RNeasy kits with DNase treatment, or similar.
  • Metagenomic DNA samples: we recommend a physical lysis method such as bead beating to preserve community diversity.
  • Long read sequencing: we strongly recommend sending raw samples for us to extract nucleic acids composed of longer fragments for better service.

We recommend Qubit or other fluorescent dye-based measures of nucleic acid quantification, or Bioanalyzer/Tapestation. Nanodrop is also acceptable, though we ask that you aim to supply well above the requested 10ng/ul minimum as this technology tends to overestimate concentrations by as much as 10-fold. When Nanodrop and Qubit values are discordant this is often an indication of contaminants such as phenol or ethanol, which can sometimes interfere with NGS workflows.

It depends on your research question! Here are a few guidelines:

  • Variant calling, or mutation detection in comparison with a reference genome, typically requires a minimum of 30x mean coverage. Converting this to our offerings, multiply [desired coverage] x [genome size, in Mb]. 
  • For de novo hybrid assembly, we recommend at least 50X short reads and 30X long reads.

For genome resequencing projects, we strongly encourage including your wild-type strain along with your test samples.

Sequencing data for most sample types is available within 14 business days of receipt of your samples, if not sooner. Bioinformatics deliverables are typically available within 7 business days of receiving your sequencing data. You will receive an email confirmation of receipt and a projected delivery date for your order.

On a tight timeline? We also offer flexible expedited turnaround options – contact us for more information!

If you have a large order, contact us to ask about reduced pricing in exchange for longer turnaround time.

  • Raw sequencing files in FastQ format
  • README files with documentation of specific methods and analyses that were performed on your samples, including bioinformatic analyses when applicable.
  • Submitting your sequences to NCBI or another data repository? You can find all of the information required for submission in the README documents you received with your data.

Data delivery methods:

  1. Secure File Transfer Protocol (SFTP) is our standard option. It is fast and secure, but we understand that some of our customers are unfamiliar with the technical procedures involved in connecting to the server. So, we have worked hard to create a clear, step-by-step SFTP Data Retrieval Guide.
  2. Box.com or AWS are our secondary options and are offered to any customer who prefers these methods. Please let us know if you would like to receive your data by one of these methods and we will accommodate your needs promptly. If you want to retrieve data from our AWS bucket, please send us your 12-digit AWS Account ID so we can provide AWS CLI access to your data.

 

Your sequencing & bioinformatics results will be readily available to download for 30 days after data delivery. They will then be moved to our long term storage for 5 months, after which they will be deleted. If you need to download your results after 30 days, please contact us and we will transfer them out of long-term storage. We highly recommend that you download your results as soon as they are available!

Within your folder, you will find compressed FASTQ files for each sample (2 per sample for paired end reads), a README of methods, sample QC files, and any analysis services you requested in a separate directory.

Credit Cards

We welcome payments via all major credit cards. Please note, credit card payments are subject to a processing fee that helps us cover part of the costs charged by our card processor, Stripe.

Purchase Orders

For organizations wishing to use purchase orders, we invite you to register SeqCoast within your organization’s procurement system. For assistance with registration, please contact our finance team at finance@seqcoast.com

Wire Transfers

Wire transfers are accepted and include a surcharge of $30.00 to cover incoming bank fees. For international transfers where the institutional policy is to deduct outgoing wire fees from the payment amount, an additional $30.00 surcharge will apply.

For any further assistance or clarification on making payments, please reach out to finance@seqcoast.com.

TL;DR

16S is used to profile microbial communities. Full-length 16S sequencing (Nanopore) yields higher resolution of species and strains compared to V3/V4 sequencing (Illumina).

Long version:

16S rRNA sequencing is used to examine bacterial species diversity and abundance of a microbial community. The 16S rRNA gene is a well-established marker for bacterial species, and it consists of both conserved regions and hypervariable regions (V1-V9). Primers bind to the conserved regions, and the hypervariable regions are amplified, sequenced, and analyzed.

Because the entire 16S sequence is about 1500bp long, only a subset of the sequence can be revealed using Illumina sequencing. We use primers 341F and 806R to amplify the V3/V4 hypervariable regions, and we perform 2x300bp paired end sequencing. This sequence length allows the paired reads to overlap, maximizing accuracy. Still, Illumina sequencing often fails to resolve species or strain-level differences. If this level of detail is important for your project, use our Nanopore full-length 16S service to sequence and analyze the entire 16S gene!

Our standard protocols are designed to accommodate a broad range of biological samples, including bacteria, fungi, algae, and various cell lines, given that the recommended inputs are provided. While we strive to ensure the highest quality and compatibility, it’s important to note that we cannot guarantee flawless performance for every species or sample type due to the inherent variability in biological materials.

We offer custom processing for complex samples such as stool, soil, filters, and others that may require specialized workflows. Please contact us to get a quote on your next custom project!

We accept all samples in PCR tubes, 1.5 mL tubes, or 96-well plates.

Samples in Tubes

When sending samples in tubes, we recommend wrapping tube lids in Parafilm when possible, and packing samples with plenty of foam or bubble wrap for protection. This helps prevent tubes from popping open or becoming crushed during transit.

Samples in 96-Well Plates

For large orders, it is often easier to send 96-well plates. We recommend using PCR plates that are well-sealed with either foil seals or plastic heat-seals. We strongly suggest shipping the plates on dry ice if possible. We find that sometimes the shaking and sloshing from the shipping process defeats even the strongest plate seals, and keeping the samples frozen helps prevent well-to-well contamination.

Additionally, in your sample manifest, it is important to indicate which sample is in which well. The best way to do this is to add a column labeled “Well ID” to your sample manifest before uploading.

Sample Preparation Guidelines

Nucleic Acid Samples

Service Concentration* Volume Buffer Shipping Considerations
Short read sequencing, Illumina
>10 ng/µL
>30 µL
nuclease-free water or dilute TE (1:10)
Can be shipped at room temperature.
Long read sequencing, Oxford Nanopore
>50 ng/µL
>60 µL
nuclease-free water
Should be shipped with ice packs or on dry ice.
†Poly-A Enrichment RNAseq, Illumina
>25 ng/µL RNA
>20 µL. Please include a second aliquot (~5 µL) for sample QC.
nuclease-free water or dilute TE (1:10)
RNA should be shipped overnight on dry ice.
†Ribodepletion RNAseq, Illumina
>25 ng/µL RNA
>20 µL. Please include a second aliquot (~5 µL) for sample QC.
nuclease-free water or dilute TE (1:10)
RNA should be shipped overnight on dry ice.
16S Amplicon (Illumina or Nanopore)
>10 ng/µL
>30 µL
nuclease-free water
Can be shipped at room temperature.
Full Flow Cell Runs, Illumina
Prepared and pooled libraries with a final concentration of at least 2 nM.
RSB + Tween
Can be shipped with ice packs or on dry ice.
Full Flow Cell Runs, Nanopore
Same as long read Nanopore
nuclease-free water
Should be shipped with ice packs or on dry ice.

*We recommend quantifying by Qubit, as Nanodrop can overestimate concentrations >10-fold if contaminants are present.

We highly recommend eluting samples in nuclease-free water or diluted TE, as excessive EDTA can interfere with with library prep. Extraction contaminants (salts, phenol, polysaccharides, etc.) can cause sample failure.

†Note for RNA services: To minimize the amount of time that your RNA samples are thawed, please label your tubes with their SeqCoast tube IDs and dilute your samples to <250 ng/µL prior to shipping. 

Services that include DNA Extraction

Service Sample Type Shipping Considerations
Hybrid whole genome sequencing, Illumina/Nanopore OR Strain typing, Illumina
Frozen cell pellets (50 - 100 mg), all supernatent removed; OR Sealed, pre-grown agar plates - must have enough growth to allow us to easily collect 50 - 100 mg of colonies.*
Ship frozen cell pellets on dry ice. Ship agar plates with ice packs or at room temperature.

*We have found that ~10e9 E. coli cells is sufficient for our extraction workflows.

Looking for detailed guidance on selecting the right sequencing approach for your science? Our consultation services provide expert advice tailored to your project's needs.