The Rapid PCR Barcoding Kit provides a fast and simple method of preparing up to 12 barcoded gDNA samples for nanopore sequencing from 1-5 ng using PCR.
- Low input amounts ― 1 to 5 ng DNA
- Fast library prep ― 15 minutes of hands-on preparation time
- Multiplex up to 12 samples
Barcoding or multiplexing is useful when the amount of data required per sample is less than the total amount of data that can be generated from a single flow cell: it allows a user to pool multiple samples and sequence them together making more efficient use of the flow cell.
The Rapid PCR Barcoding Kit offers the fastest and simplest method of preparation of barcoded libraries for low quantities of gDNA (1 to 5 ng), with only approximately 15 minutes of hands-on preparation time.
At the heart of the kit is a transposase which simultaneously cleaves template molecules in each sample and attaches tags, which contain primer binding sites, to the cleaved ends. The kit contains 12 primers which are then used to amplify each sample: each primer contains a barcode and 5’ tag which facilitates the ligase-free attachment of Rapid Sequencing Adapters. Amplified and barcoded samples are then pooled together and Rapid Sequencing Adapters are added to the pooled mix.
This kit is optimized to offer a simple workflow for those with a limited amount of input material. It has been observed that the fragment length produced after PCR is largely independent of the input fragment length, with the PCR products producing a peak at around 2 kb on an Agilent Bioanalyzer. This corresponds to a read-length of ~2 kb in nanopore sequencing. The length limit is due to the method of library preparation, rather than the processivity limit of the DNA polymerase.
Deconvolution of barcoded sequencing data is supported by the MinKNOW software, the stand-alone Guppy basecaller, and EPI2ME, all of which classify the barcode sequence and sort reads into corresponding folders.
If larger gDNA reads are required, see the PCR Barcoding Kit.
Sample multiplexing is ≤12.
Informacja o dostawie: Flow cells and kits are shipped together at 2 to 8 °C. Upon receipt, please place the product in a long-term storage location at ‒20 °C. Oxford Nanopore Technologies deem the useful life of the product to be three months from receipt by the customer.
Uwaga: Oxford Nanopore Technologies products are currently for research use only.