Detection of small interfering RNAs by rolling circle amplification

CHED 204

Ni Li, ni.li@email.ucr.edu1, Sonja Kress, sonjakress@yahoo.com2, and Wenwan Zhong, wenwan.zhong@ucr.edu1. (1) Department of Chemistry, University of California at Riverside, Riverside, CA 92521, (2) Leibniz University, Welfengarten 1, Hannover, 30167, Germany
Studying the expression profiles of siRNAs in plants is very important for understanding their functions and relevance in the host's viral defense system. Consequently there is great demand for a simple, reliable and sensitive siRNA detection method. Here we present a method based on padlock probes and rolling circle amplification to quantitatively analyse siRNAs in limited amount of total RNA. The presence of a specific siRNA strand annealed to a corresponding ssDNA template leads to a ligation with the termini of the padlock probe and therefore forms a circular DNA, which can be amplified isothermally. This process creates an ssDNA product containing multiple copies of the siRNA sequence. Additionally a sequence for a DNAzyme, which acts as a catalytic unit, will be inserted into the repeated sequence. Thereby multiple enzymes can be generated and detected by a specific enzyme reaction to further amplify the signal from a specific siRNA.
 

Undergraduate Research Poster Session
2:30 PM-4:30 PM, Monday, August 20, 2007 BCEC -- Exhibit Hall - B2, Poster

Division of Chemical Education

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007