OCTOPUS (Optimized Cloning Through Open-source Pipelined Unabridged Sequencing), is a light-weight, cost-effective, and robust method for full-plasmid sequence verification using next-generation sequencing. Importantly, OCTOPUS can be performed at scale with common lab equipment and uses crude E. coli lysate (rather than purified DNA) as the input. This reduces up-front capital investment by eliminating the need for expensive nano-volume liquid handlers and automated plasmid purification pipelines. Using OCTOPUS, a single molecular biology researcher can sequence verify 96-1152 colonies in three days for ~$5 a sample.
How OCTOPUS Works
OCTOPUS is divided into six main steps.
- colony picking and overnight growth
- lysate and glycerol stock generation
- iGenomX Riptide kit library preparation
- Illumina sequencing
- turn-key data analysis
- pick perfect sequences
Why we Love It
We've used OCTOPUS to sequence ~10,000 plasmids in a 6 month period.
Please feel free to open an issue or pull request.
This project is licensed under the Apache 2.0 License - see the LICENSE file for details. Additional licensing information: