quantification of the expression of pre-miRNA and miRNA in plants - (Mar/23/2010 )
I would like to check the expression level of specific miRNA precursor and mature miRNA in Arabidopsis. Does anyone have protocol/experience/suggestion to do the job? and how to design the primers? Thanks a lot!
one traditional method is the extracting RNA adding polA to the 3' ends by polA polymerase then making cDNA with a universal odT primer(an oligo dT with a universal flanking adaptor at 5prime end with no homology to plant genome or RNA).you just design a forward primer for precursor(the exact seq of precursor) and a reverse primer complementary to the adaptor region
search key words like miRNA expression,polA polymerase... for articles, i do not have their titles in my mind