Plasmid sequencing
Full-length plasmid sequencing for $20 CAD per plasmid
Flow Genomics is partnering with the London Regional Genomics Center to provide low-cost full plasmid sequencing. This service is optimized for plasmid sizes from 2-30 kb. For a quote on larger plasmids, please contact info@flowgenomics.com.
Sample requirements
Please submit at least 10 uL diluted to 30 ng/uL in water or low-EDTA buffer. We recommend using water when possible.
Recommended DNA quality
- 10 uL of 30 ng/uL in water (by Qubit)
- A260/280 of 1.8 (by Nanodrop)
- A260/230 > 1.8 (by Nanodrop)
Lower quality samples often can be sequenced, but cannot be guaranteed.
Qubit quantification is more accurate than Nanodrop, which tends to over-estimate the concentration and may lead to failures. No amplification steps are performed for plasmid sequencing - everything that is in the sample is sequenced, even if contaminated with genomic DNA. Low-quality samples (low purity, contaminated with genomic DNA or RNA, degraded plasmids) will yield poor data. No refunds will be issued for failed samples when DNA provided is poor quality.
Want to set up a drop box at your institution? Email us!
Sample submission
Please send plasmids in 8-strip tubes only.
Please courier or bring samples to:
London Regional Genomics Centre
Robarts Research Institute
Room 4212
1151 Richmond Street North
London, Ontario, N6A 5B7
(519)931-5777 Ext. 24074
Please ship at ambient temperature.
What we send you
We send the following files:
- FASTA of each plasmid assembled
- report that can be opened in browser containing quality information and annotations
Sequencing quality
We sequence plasmids using the Oxford Nanopore Technologies platform. After sequencing to a high read depth, Q50 full plasmid sequences are typically achieved (i.e., 1 error in 10 000). The reminaing errors are typically at homopolymer regions. Here's what Oxford Nanopore says about their quality: https://nanoporetech.com/accuracy
Annotation
Each plasmid is annotated as below (pGem) using pLannotate:
Quality information
Sequencing quality of all reads for each plasmid is plotted for troubleshooting purposes. Ideally, most of the sequencing reads should be full-length plasmids as below (e.g., ~3kb reads is the full pGem plasmid).
Questions? Feedback? Collaborate? Reach out to info@flowgenomics.com.