[커스텀 프로토콜] Nanopore Sequencing Kit 사용자 프로토콜 모음

Nanopore Sequencing Kit Protocols

These protocols use information from Oxford Nanopore Technologies' protocols and have been written by me; I release them to the world under a CC-BY-SA license. The source .Rmd text for all versions can be made available; please let me (David Eccles) know if that would be useful to you.

If you find these protocols useful, please consider buying me a ko-fi. Or better yet, think of me when you have a little bioinformatics project that I could help out with. More details about what I do can be found here:

https://www.gringene.org/services.html

PCR cDNA Barcoding Kit Protocol

The PCR cDNA Kit is used for high-accuracy quantification of cDNA from extracted RNA. If sample amounts are abundant, the number of cDNA cycles can be reduced in order to lessen the impact of PCR-related artefacts.

• Version 1.1.6 - added GrinGene logo
• Version 1.1.5 - [re] unified all loading protocols to RBK protocol
• Version 1.1.4 - Added adapter information to introduction
• Version 1.1.3 - added rapid adapter references
• Version 1.1.2 - added quick reference flow cell loading protocols
• Version 1.1.1 - fixed up LongAmp mention for PCR master mix
• Version 1.1.0 - updated to a non-ligation cDNA protocol

Rapid Barcoding Kit Protocol

The Rapid Barcoding Kit is a cheap, fast protocol; great for plasmids, amplicons, and any situations where there is lots of high-quality DNA. Because the transposase fragments DNA, it can be used on circular DNA (e.g. plasmids, mitochondria) without any additional linearisation.

• Version 2.2.3 - Updated to be more clear about flongle loading
• Version 2.2.2 - added GrinGene logo
• Version 2.2.1 - clarified some steps, removed tube transfer at end of bead purification
• Version 2.2.0 - Updated to ONT's new protocol (1.5 μl RBXX, 1X XP Beads, elution in 30 μl EB/TBS). Adjustment of SB/DNA amounts for Flongle loading
• Version 2.1.0 - added quick summary
• Version 2.0.0 - updated to V14 kit

Ligation Sequencing Kit Protocol

The Ligation Sequencing Kit is a stable / robust protocol that delivers excellent sequencing yields; great for large genome sequencing, or situations where getting the highest yield from a sample is important. Note that because this is a ligation protocol, there is a higher chance of ligation-associated artefacts (chimeric reads) being generated during the sample preparation process.

• Version 1.0.2 - added GrinGene logo
• Version 1.0.1 - [re] unified all loading protocols to RBK protocol
• Version 1.0.0 - V14 kit with quick summary

Rapid PCR Barcoding Kit Protocol

The Rapid PCR Barcoding Kit is a moderately cheap and fast protocol that works great for metagenomic applications, and any situations where sample amounts are low (less than 5ng). It produces read lengths of about 1-3kb, which is usually fine for microbial assembly, but can be too short for large genome assembly

• Version 1.0.0 - updated to V14 kit and added GrinGene logo

Flongle Flow Cell Loading Protocol

• Version 2.2.0 - added GrinGene logo, and changed loading volumes to 30μl to be consistent with brief protocols

https://www.gringene.org/nanopore/protocols/