Jul 31, 2025
Version 1
Library construction for ONT long read cDNA V.1
- corinne blugeon1
- 1genomiqueENS
- genomiqueENS
Protocol Citation: corinne blugeon 2025. Library construction for ONT long read cDNA. protocols.io https://dx.doi.org/10.17504/protocols.io.4r3l21oepg1y/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 31, 2025
Last Modified: July 31, 2025
Protocol Integer ID: 223791
Keywords: RNASeq, ONT, long read, read cdna library construction, length rna, rna, library construction for ont, cdna, cdna this protocol, using ont technology, ont technology, ont, phred score, mammalian sample, library construction, successful retrieval, average from mammalian sample
Abstract
This protocol details the optimization of long-read cDNA library construction using ONT technology. This method allows for the successful retrieval of at least 2kb on average from mammalian samples. It has been adapted to ONT's latest chemistry 14. Phred scores above 20 can be achieved with near-full-length RNA.
Troubleshooting
Protocol references
Takara Bio USA, Inc. SMART-Seq‱ mRNA
Oxford nanopre technologies ; PCR Expansion ;EXP-PCA001
Oxford nanopre technologies ; cDNA-PCR Sequencing Kit V14 SQK-PCS114