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Native Barcoding Expansion 1-12 and 13-24 features
These kits are recommended for users who:
- wish to multiplex samples to reduce price per sample
- need a relatively quick method of multiplexing, avoiding the need of further PCR
- want to optimise their sequencing experiment for throughput
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Introduction to the Native Barcoding protocol
This protocol describes how to carry out native barcoding of amplicons using the Native Barcoding Expansion 1-12 (EXP-NBD104) and 13-24 (EXP-NBD114), in conjunction with the Ligation Sequencing Kit (SQK-LSK109). There are 24 unique barcodes if using both expansion kits, allowing the user to pool up to 24 different samples in one sequencing experiment. It is highly 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:
- 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 runPrepare your library
You will need to:
- Prepare the DNA ends for adapter attachment
- Attach Native barcodes supplied in the kit to the DNA ends
- Attach sequencing adapters supplied in the kit to the DNA ends
- Prime the flow cell, and load your DNA library into the flow cellSequencing
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 barcoding workflow