Single-ribonucleotide repair-mediated ligation-dependent cycling signal amplification for sensitive and specific detection of DNA methyltransferase

Abstract

Specific and sensitive detection of DNA MTase activity can be achieved by a single-ribonucleotide repair-mediated ligation-dependent cycling signal amplification approach.

A single quantum dot-based nanosensor with multilayer of multiple acceptors for ultrasensitive detection of human alkyladenine DNA glycosylase

“…We designed an AP probe with an AP site (green + purple color, Scheme 1) at the 23rd base from the 5′ end. The AP probe is paired with the trigger (pink color, Scheme 1) generated by the cleavage of the hairpin probe for the initiation of isothermal strand displacement amplification (SDA)41,42 in the presence of APE1. In addition, we designed a capture probe (yellow color, Scheme 1) which can hybridize with the primer (green color, Scheme 1) for the initiation of DNA polymerase-assisted amplification.…”

Rolling circle amplification-driven encoding of different fluorescent molecules for simultaneous detection of multiple DNA repair enzymes at the single-molecule level

“…35 Isothermal amplication of nucleic acids (e.g., rolling circle amplication (RCA)) as an alternative amplication technique enables rapid and effective amplication at constant temperature. [35][36][37][38][39][40][41][42][43][44] RCA may generate very long single-stranded DNAs (ssDNAs) with tandem repeats, and it can achieve approximately 10 3 -fold amplication within 1 h. 35,[45][46][47] The visualization and analysis of RCA products usually use either SYBR green I as the uorescence label 45 or the uorescence-labeled oligonucleotide probe. 46 SYBR Green I is a DNA intercalating dye that binds dsDNA, 45 but it has several limitations including the concentration-dependent inhibition of PCR, preferential binding to the GC-rich sequences, the promotion of nonspecic amplication, and the detection of only a single type of target due to the use of a single uorophore, false positive signals due to its binding to any dsDNAs including nonspecic dsDNA sequences.…”

An integrated-molecular-beacon based multiple exponential strand displacement amplification strategy for ultrasensitive detection of DNA methyltransferase activity

“…For example, Yuan's group reported a fluorescence method based on exonuclease III (Exo III)-assisted isothermal cycling signal amplification;31 Zhang's group reported a surface-enhanced Raman scattering method based on strand displacement amplification (SDA) strategy;32 Zhu's group reported a chemiluminescence strategy combining hybridized chain reaction (HCR) and rolling circle amplification (RCA);33 and recently, Zhang's group reported an RNase HII-assisted single-ribonucleotide repair-mediated ligation-dependent cycling signal amplification method 34. Nevertheless, the complicated signal amplification process usually requires a sophisticated design of the DNA template and probe sequences 34. And numerous DNA oligos, which have to be involved in the system to assist the reaction, inevitably generate high background because of the nonspecific amplification 33.…”