Journal
METHODS AND PROTOCOLS
Volume 6, Issue 2, Pages -Publisher
MDPI
DOI: 10.3390/mps6020031
Keywords
amplification-free protocol; direct cDNA sequencing; high-throughput sequencing; MinION; Oxford Nanopore Technologies((R)); poly(A) RNA; Solanaceae; Solanum betaceum; transcriptome; woody plant
Categories
Ask authors/readers for more resources
Long-read sequencing methods enable comprehensive analysis of transcriptomes, improving gene annotation and gene expression studies. However, woody plant tissues pose challenges for cDNA library preparation, hindering cutting-edge sequencing analyses.
Long-read sequencing methods allow a comprehensive analysis of transcriptomes in identifying full-length transcripts. This revolutionary method represents a considerable breakthrough for non-model species since it allows enhanced gene annotation and gene expression studies when compared to former sequencing methods. However, woody plant tissues are challenging to the successful preparation of cDNA libraries, thus, impairing further cutting-edge sequencing analyses. Here, a detailed protocol for preparing cDNA libraries suitable for high throughput RNA sequencing using Oxford Nanopore Technologies((R)) is described. This method was used to prepare eight barcoded cDNA libraries from two Solanum betaceum cell lines: one with compact morphology and embryogenic competency (EC) and another with friable and non-embryogenic (NEC). The libraries were successfully sequenced, and data quality assessment showed high mean quality scores. Using this method, long-read sequencing will allow a comprehensive analysis of plant transcriptomes.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available