TY  - JOUR
TI  - Circular RT-PCR Assay Using Arabidopsis Samples
AU  - Hang, Runlai 
AU  - Deng, Xian 
AU  - Liu, Chunyan 
AU  - Mo, Beixin 
AU  - Cao, Xiaofeng 
VL  - 5
IS  - 14
PY  - 2015
DA  - 2015/07/20
SP  - e1533
C1  - Bio-protocol 2015;5:e1533
DO  - 10.21769/BioProtoc.1533
UR  - https://doi.org/10.21769/BioProtoc.1533
AB  - Post-transcriptional processing is critical for RNA biogenesis, in which conventional functional RNA transcripts are generated, such as messenger RNAs (mRNAs), transfer RNAs (tRNAs) and ribosomal RNAs (rRNAs) for translation as well as emerging non-coding RNAs with known or unknown regulatory functions. To determine the precise termini of an RNA molecule during or after processing, the primer extension and Rapid Amplification of cDNA Ends (RACE) methods have been routinely utilized for the precise mapping of 5’ or 3’ ends. Different from these assays, which are designed to detect only one end of a specific target RNA at a time, circular Reverse Transcription-Polymerase Chain Reaction (cRT-PCR) is able to simultaneously determine both the 5’ and 3’ ends of the target RNA. In Arabidopsis thaliana, cRT-PCR has been wildly applied to identify both the 5’ and 3’ extremities of the ribosomal RNA precursors, or to assess the length or post-transcriptional extensions at the 3’ end of a matured mRNA. In this protocol, we summarize and present a detailed procedure of the cRT-PCR assay in Arabidopsis thaliana, which is also successfully used in our previously published work (Hang et al., 2014).
KW  - Circular RT-PCR
KW  - Post-transcriptional processing
KW  - Pre-rRNA processing
KW  - Ribosome biogenesis
KW  - PRMT3
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False