Significant innovations in next-generation sequencing techniques and bioinformatics tools have impacted our appreciation and understanding of RNA. Practical RNASeq applications have evolved in conjunction with sequence technology and bioinformatic tool advances. In this review, we explained various computational resources, tools, and bioinformatics analyses advancement for small and large non-coding RNAs. These include non-coding RNAs (ncRNAs) such as piwi, micro, circular, and long ncRNAs. In addition, this article discusses future challenges, single-cell level sequencing for non-coding RNAs, and advantages of using long-read sequencing to annotate lncRNAs.