CHMI owns and operates its own MiniSeq. This instrument is ideal for a bacterial genome sequencing, PCR panel assays, and low-coverage QC runs prior to larger NovaSeq runs.
- Reference material
- Thawing the Kit
- Denaturation and Dilution
- Local Run Manager
- Loading the Sequencer
- Sequencing by Synthesis: 5 minute YouTube video
- MiniSeq system guide
- MiniSeq denature and dilute guide
- MiniSeq support site
- MiniSeq training videos
Thawing the Kit
Denaturation and Dilution
The following protocol is based on the MiniSeq Denature and Dilute Guide (linked above), with few adjustments. The biggest change is that we do not vortex the libraries after denaturation.
Before proceeding to these steps, make sure your flow cell is out at room temperature and your reagent cartridge is thawing. In addition to these sequencing reagents, you will need your pool, 20pM denatured PhiX, 1M Tris-HCl pH 8, 1N NaOH, MilliQ water, RSB, and thawed Hybridization Buffer.
|Desired Loading Concentration pM||ul Hyb Buffer||ul 5pM Library Pool||ul 20pM PhiX for 1% Spike-In||ul 20pM PhiX for 5% Spike-In||ul 20pM PhiX for 20% Spike-In|
Local Run Manager
Loading the Sequencer
Before loading, make sure your run has been programmed in Local Run Manager (on the MiniSeq) or another sample sheet template.