Crispr rna crrna

All reactions involved primers, ssDNA-reporter (FAM-TTATT-BHQ1), and CRISPR RNA (crRNA) synthesized by Sangon Biotech (Shanghai, China). EngenLba Cas12a, nuclease-free water, and 10 × NEbuffer were purchased from New England Biolabs (Ipswich, MA, United States). The TwistAmp Basic kit (#TABAS03KIT) for RPA was purchased from TwistD× Limited (Maidenhead, United Kingdom). The PCR Master Mix was purchased from Takara (Dalian, China). The Quantitative Real-time PCR (qPCR) Master Mix was purchased from YEASEN (Shanghai, China). The pUC57 recombinant plasmids containing vacA, cagA, and 16SrDNA genes of H. pylori were transformed by E. coli, and the above process was done by Sangon Biotech (Shanghai, China). Genomic DNA Extraction Kits were purchased from TIANGEN Biotech (Beijing, China).
Clinical saliva samples were provided by the Tang du Hospital of Air Force Military Medical University (Xi’an, China). The study was approved by the Air Force Military Medical University Research Ethics Review Board. A written informed (verbal) consent was obtained from all patients. Saliva samples were collected from all volunteers after undergoing ¹³C-UBT.

Monodispersed Magnetic Streptavidin Microspheres (MBs) were purchased from Tianjin Bessel Chromatography Technology Development Center (Tianjin, China), with a solid content of 0.5% (w/v) and a particle size of 300 nm. ZEN, Ochratoxin a (OTA), Fumonisin (FB1), Aflatoxin B1 (AFB1) and Aflatoxin M1 (AFM1) were bought form Meizheng Testing Technology Co., Ltd. (Beijing, China). EnGen LbaCas12a from Lachnospiraceae bacterium ND2006 was purchased from New England Biolabs (Ipswich, MA, USA). CRISPR RNA (crRNA) and other ssDNAs were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). Sequence details are given in Table S1. The buffer solution 1 was used as a reaction solution for the streptavidin-modified magnetic beads and DNA. The CutSmart Buffer and NEBuffer™ 2.1 was from New England Biolabs. Buffer components are visible in Table S2. PBS, agarose, 50 × TAE, 5 × TBE, EDTA, 30% acrylamide solution (ACR-BIS, 29:1), tetramethylethylenediamine (TEMED), PAGE coagulant, ammonium persulfate (APS)⁺, 6× loading buffer and DNA Marker were purchased from Sangon Biotechnology Co., Ltd. (Shanghai, China). RNase-free water was employed as the solvent throughout the experiment. All chemical reagents were of analytical grade.

The RNA standards for SARS-CoV-2, SARS-CoV, Parainfluenza virus, Influenza virus A (FluA), and Influenza virus A (FluB) were synthesized by the Shanghai Institute of Measurement and Testing Technology. All primers, CRISPR RNA (crRNA), and the ssDNA reporter (FAM-TTATT-BHQ1) were synthesized by the Sangon Biotech Co. Ltd., Shanghai, China. EnGen^® Lba Cas12a was purchased from New England Biolabs (Ipswich, MA, UK). The RNase inhibitor was purchased from Takara and the RPA amplification kit was purchased from Leshang Biotechnology (Wuxi) Co., LTD., China.