Challenges for Accurate Quantification of RNA

Ribonucleic acid (RNA) quantification is an essential technique in biology. There has been remarkable progress RNA techniques over the recent years; however, specificity of these to quantify a very small amount doubtful because factors which can inhibit precise quantification. To develop that leads most sensitive quantification, problems must be overcome. In this article, we first review RNA: quality RNA, secondary structure efficiency enzyme reaction, annealing conditions, limitations experimental protocol and equipment, detection sensitivity equipment. Next, discuss possible methods contribute factors: control focused on target degradation, isothermal amplification, for avoiding amplification errors, RNase H-dependent PCR, targeting using fluorescent-labeled probe, padlock bridged/locked nucleic (BNA/LNA) peptide (PNA), clustered regularly interspaced short palindromic repeat (CRISPR) system. One goals development ultrasensitive absolute RNA. Here, used type