Selected article for: "background fluorescence and fluorescence signal"
Author: Lu Guo; Xuehan Sun; Xinge Wang; Chen Liang; Haiping Jiang; Qingqin Gao; Moyu Dai; Bin Qu; Sen Fang; Yihuan Mao; Yangcan Chen; Guihai Feng; Qi Gu; Liu Wang; Ruiqi Rachel Wang; Qi Zhou; Wei Li
Title: SARS-CoV-2 detection with CRISPR diagnostics
  • Document date: 2020_4_11
    • ID: 2una9767_7
    Snippet: We found out that increasing the sgRNA concentration by 3 folds not only enhances the fluorescence signal and the signal-to-background ratio, but also increases the rate of reaction Given that the average viral load in the plasma of SARS patients ranged from less than 1 to about 1000 copies per microliter [9] , or 1 × 10 3 -1 × 10 6 copies/mL, nucleic acid amplification techniques are needed to produce sufficient DNA for CRISPR-based DNA detect.....
    Document: We found out that increasing the sgRNA concentration by 3 folds not only enhances the fluorescence signal and the signal-to-background ratio, but also increases the rate of reaction Given that the average viral load in the plasma of SARS patients ranged from less than 1 to about 1000 copies per microliter [9] , or 1 × 10 3 -1 × 10 6 copies/mL, nucleic acid amplification techniques are needed to produce sufficient DNA for CRISPR-based DNA detection methods. Recombinase-aided amplification (RAA) can amplify substrates 10 10 times at most (from aM to 10 nM) within 10-30 minutes at constant temperature between 37 o C to 42 o C, complementing the needs of CRISPR-based detection. Thus, we designed and screened RAA primers that matched our previously optimized sgRNA-3 (Supplementary information, Fig. S4a ). Based on our screens, we found that by using the best primer pairs together with sgRNA-3, we can detect SARS-CoV-2 RNA in samples containing 5 × 10 3 copies/mL ( Fig. 1c and Supplementary information, Fig. S4b-c) .

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