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Rapid Barcoding Kit features
This kit is recommended for users who:
- Wish to multiplex samples to reduce price per sample
- Need a PCR-free method of multiplexing to preserve additional information such as base modifications
- Require a short preparation time
- Have limited access to laboratory equipment
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Introduction to the Rapid Barcoding Sequencing Kit
This protocol describes how to carry out rapid barcoding of plasmid DNA using the Rapid Barcoding Sequencing Kit (SQK-RBK004).
Steps in the sequencing workflow:
Prepare for your experiment
You will need to:
- Extract your plasmid 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 runLibrary preparation
You will need to:
- Tagment your DNA using the Fragmentation Mix RB in the kit; this simultaneously attaches a pair of barcodes to the fragments
- Pool the barcoded samples
- Attach sequencing adapters supplied in the kit to the DNA ends
- Prime the flow cell, and load your DNA library into the flow cellSequencing 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 the analysis workflow for clone validation