Full-Length Transcriptome Analysis Using a Bias-Free cDNA Library Prepared with the Vector-Capping Method
Full-length complementary DNAs (cDNAs) are an essential resource for functional genomics. Recently, we have developed a simple and efficient method for preparing a full-length cDNA library from a small amount of total RNA, named the “vector-capping” method. The biggest advantage of this method is that the intactness of the cDNA can be assured by the presence of dG at the 5′ end of the full-length cDNA. Furthermore, the cDNA library represents the mRNA population in the cell owing to a bias-free procedure. In this chapter, we describe not only the protocol for preparing the library but also the points for analyzing the 5′-end sequence of the obtained cDNA.
Key wordsTranscriptome Full-length cDNA Vector primer Vector-capping method Expression profile Transcriptional start site Splicing isoform Long-sized cDNA
This work was supported by a grant from the Ministry of Health, Labor, and Welfare of Japan.