Thank you for your interest in the NWGC!
Below is the step-by-step process for getting your already constructed libraries sequenced. Please review these steps carefully and if you have any questions, please don’t hesitate to ask.
1.) Request a Quote
To request a quote, please email nwgc@uw.edu with the following information:
Investigator Name
Phone Number
Email Address
Request Information
*Request Information should include as much detail as possible about your project requirements. For guidance, please see the Getting Started pages under your project type for more information.
The NWGC will send you a quote based on your project specifications. Do not send any samples before reviewing the quote and returning the signed quote or purchase order to the NWGC.
2.) Sample Submission Information
Once a quote is received and signed, please email the NWGC Sequencing Team at nwgcseq@uw.edu to receive the Sample Submission Form. Please fully fill out the form and return it to nwgcseq@uw.edu prior to sample drop-off or delivery. Please also send all the barcode information for your library(s).
**We use the information provided to load the sequencing instruments and generate FASTQs. Please make sure it is as accurate as possible.
Please follow the guidelines for submission below:
Details on Required Sample Information:
- Please only submit samples in 1.5mL – 2.0mL tubes with snap-caps or screw caps!
- Please avoid naming samples 1, 2, 3 etc. and include a unique identifier in order for us to keep better track of your sample.
- The molarity of your final submission, whether that be individual libraries or a pool. Please use something more accurate than a Nanodrop, like qPCR for example.
- The read length for your sequencing run. (PE25 = 50 cycles, SE50 = 50 cycles).
- The number of base pairs in your index sequence. (ie. +8,+8+8,+10,+10+10)
- The actual index sequences, listed in the forward orientation, and what samples they correspond to. We use this to demultiplex your libraries. Please list on a new page or attach a separate file.
- The required primer type. If using custom primers (non-Illumina),please provide them at 100uM.
- The type of molecule (RNA, genome) or specialized prep (ChIP-seq, ATAC-seq) we will be sequencing.
4.) QC Results
All samples will be run on the bioanalyzer and qubit. QC results will be emailed to the investigator prior to sequencing.
5.) Data Delivery/Globus Access
Globus is a service provider that manages login credentials and coordinates transfers between organizations. Please visit the Data Delivery section for more information.
Once your run is complete, we will send you an email with a link to your specific Globus folder where the run file is located.
Supplemental Info:
- Below are the minimum volume requirements, based on our minimum molarity accepted per sequencing platform:
Sequencing Platform | Minimum Molarity | Minimum Volume |
Illumina® NovaSeq S4 | 5nM | 130uL |
Illumina® NovaSeq S2 | 5nM | 55uL |
Illumina® NovaSeq SP/S1 | 5nM | 35uL |
Illumina® NovaSeq X Plus 1.5B | 5nM | 20uL |
Illumina® NovaSeq X Plus 10B | 5nM | 60uL |
Illumina® NovaSeq X Plus 25B | 5nM | 100uL |
Illumina® NextSeq 500 MO or HO | 5nM | 10uL |
Illumina® MiSeq | 4nM | 20uL |
Element AVITI™ Cloudbreak Freestyle | 5nM | 20uL |
Long-Read Sequencing Platform | Minimum Concentration | Minimum order |
PacBio Revio™ | 20-60ng/ul in 28ul (concentration varies based on insert size) | SMRT™ Cell |
Oxford Nanopore Technologies® | 2-10ug (concentration varies based on prep type and coverage needed) | Flow cell |
- QC is included in the pricing and will be run in-house using the Qubit and Bioanalyzer for every submission. If you have any Bioanalyzer traces and/or MiSeq data available, please include with your sample submission.
- Our QC data will be used to verify the molarity of your submission. The Bioanalyzer also examines possible adapter peaks, mean insert size, and overall sample quality. The Qubit gives concentration (ng/uL) and is taken in tandem with the BA insert size to yield molarity (nM). (ng/uL)*(1,000,000) / (insert size)*(660) = nM
- Please include your sample barcode list with sample submission form prior to the sample run. For index orientation, please list the forward sequence and do not try to orient them based on instrument chemistry. We will do that when we create your FASTQ.
- FASTQ is the standard data delivery type. If you require an alternative, please inquire before submitting as an additional charge may occur.
- Once emailed that your data is ready on Globus, you will have 120 days to access it before it is deleted indefinitely.
- Notes for 10X Libraries:
- 10X Genomics Sequencing – We offer two pipelines: CellRanger version 7.0.1 and CellRanger-ATAC version 1.2.0
- For more information on 10X Genomics Sequencing, please visit their website.