Abstract
Laser microdissection (LMD) is a powerful tool for tissues or single-cell analyses. However, the difficulty of preparing cryosections and total RNA from plants constrains the technology to be unpopular in plant research. Although T7 RNA polymerase-based in vitro transcription protocol has been widely used for amplification from a hint of total RNA, the average length of cDNA becomes one-forth in 2 rounds of amplification. Thus, we employed the SMART protocol for preparing cDNA libraries from about 0.5 ng of total RNA that was extracted from pith of Arabidopsis stem by LMD, and obtained 3,697 Expressed Tag Sequences (ESTs). BLAST search suggested that about 30% of cDNA clones were full-length. As 70% of the ESTs were singletons that indicates the protocol had sufficient quality for further transcriptome analyses. We will also present the comparative study of these protocols when applied to microarray analysis.
