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Ligation Sequencing Kit XL V14 features
This kit is recommended for users who:
- Would like to process multiple samples simultaneously, either with a multichannel pipette or a liquid-handling robot
- Would like to achieve raw read sequencing modal accuracy of Q20+ (99%) or above
- Require control over read length
- Would like to utilise upstream processes such as size selection, whole genome amplification, or enrichment for long reads
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Introduction to the automated Ligation Sequencing protocol for DNA
This protocol describes how to carry out sequencing of a DNA sample using the Ligation Sequencing Kit XL (SQK-LSK114-XL).
We have developed this automated protocol on the Hamilton NGS STAR 96 liquid handling robot. The majority of the process is automated with minimal hands-on time which is required for sample quantification and deck re-loading.
It is highly recommended that a Lambda control experiment is completed first to become familiar with the technology.
To efficiently load multiple PromethION Flow Cells, we recommend using the Loading multiple PromethION Flow Cells protocol as a guideline.
Steps in the sequencing workflow:
Prepare for your experiment
You will need to:- Extract your DNA and check its length, quantity and purity. The quality checks performed during the protocol are essential in ensuring experimental success.
- Ensure you have your sequencing kit, the correct equipment, primed liquid-handling robot and third-party reagents
- Download the software for acquiring and analysing your data
- Check your flow cells to ensure they have enough pores for a good sequencing run
Library preparation
You will need to:- Repair the DNA and prepare the DNA ends for adapter attachment
- Attach sequencing adapters supplied in the kit to the DNA ends
- Prime the flow cell and load your DNA library into the flow cell
Sequencing and analysis
You will need to:- Start a sequencing run using the MinKNOW software which will collect raw data from the device and convert it into basecalled reads