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Terminal deoxynucleotidyl transferase combined CRISPR-Cas12a amplification strategy for ultrasensitive detection of uracil-DNA glycosylase with zero background

Biosens Bioelectron. 2021-01; 
Yi-Chen Du , Si-Yuan Wang , Ya-Xin Wang , Jia-Yi Ma, Dong-Xia Wang, An-Na Tang, De-Ming Kong
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Custom DNA/RNA Oligos RNA sequence was synthesized and HPLC-purified by Genscript Biotech (Nanjing, China) Get A Quote

摘要

A simple and highly sensitive biosensing strategy was reported by cascading terminal deoxynucleotidyl transferase (TdT)-catalyzed substrate extension and CRISPR-Cas12a -catalyzed short-stranded DNA probe cleavage. Such a strategy, which is named as TdT-combined CRISPR-Cas12a amplification, gives excellent signal amplification capability due to the synergy of two amplification steps, and thus shows great promise in the design of various biosensors. Based on this strategy, two representative biosensors were developed by simply adjusting the DNA substrate design. High signal amplification efficiency and nearly zero background endowed the biosensors with extraordinary high sensitivity. By utilizing these two biosen... More

关键词

Amplification; CRISPR-Cas12a; Uracil-DNA glycosylase (UDG); Zero background.