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Ligation Sequencing Kit XL 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 Ligation Sequencing Kit XL (SQK-LSK112-XL) protocol
This protocol describes how to carry out sequencing of multiple DNA samples simultaneously using the Ligation Sequencing Kit XL (SQK-LSK112-XL). It is recommended that a Lambda control experiment is completed first to become familiar with the technology.
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 and third-party reagents
- Download the software for acquiring and analysing your data
- Check your flow cell to ensure it has enough pores for a good sequencing run
Library preparationYou 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
- Start the EPI2ME software and select a workflow for further analysis (this step is optional)