AI Article Synopsis
- The study introduces a protocol that enhances long-read sequencing of specific areas in potato cyst nematodes without needing DNA amplification, utilizing a Cas9-based Nanopore technique.
- It outlines the necessary steps for designing precise guide RNAs, extracting high-quality DNA from the nematodes, and preparing the genomic DNA for sequencing.
- The protocol includes how to use the Cas9-guide RNA complex for targeted DNA cutting, followed by preparing the samples for Nanopore sequencing, with additional details referenced from Sonawala et al.
Article Abstract
We present a protocol to achieve a higher depth of long-read sequencing of region(s) of interest in potato cyst nematodes without amplification using a Cas9-based Nanopore enrichment approach. We describe steps for designing high-fidelity guide RNAs to be used with Cas9 nuclease, extracting high-molecular-weight DNA from the nematodes, and dephosphorylating genomic DNA ends. We then detail procedures for using Cas9-guide RNA complex to make targeted cleavage of the region of interest followed by a Nanopore library preparation. For complete details on the use and execution of this protocol, please refer to Sonawala et al..
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11566856 | PMC |
| http://dx.doi.org/10.1016/j.xpro.2024.103427 | DOI Listing |
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