Abstract
Introduction Recent development in oligonucleotide synthesis and DNA sequencing technologies have facilitated advancements for application of the human genome sequencing project into the clinics and routine research thus advance human health and our understanding of human biology. Examples are exome sequencing (1), CRISPR genome editing (2) and MPRA (3,4). Aside from analysis of liquid biopsies, sample-derived cell lines, tissue imaging and investigation of molecular events in situ (5, 6, 7) also provides a means to retrieve biological information however argument on specificity from the secondary labels. Present study built on previous work on utilizing several molecular biology tools, such as restriction enzymes (9) and rolling circle amplification (RCA) for enhancing signal-to-noise ratio in a detection assay (8) – a novel probe-type 2fold probe is developed to generate a single strand circular reporter molecule upon target recognition and multi-step enzymatic conversion. We specifically investigated probe performance by varying a 9-nt sequence within the stem structure at single nucleotide resolution and in library fashion.
Competing Interest Statement
The authors have declared no competing interest.