Author: Yu, Fei; Qiu, Ting; Zeng, Ying; Wang, Yiyin; Zheng, Shufa; Chen, Xiao; Chen, Yu
Title: Comparative Evaluation of Three Preprocessing Methods for Extraction and Detection of Influenza A Virus Nucleic Acids from Sputum Document date: 2018_3_2
ID: 0kt9b1ww_11
Snippet: RNA was extracted simultaneously everyday by using the Automated Nucleic Acid Extraction System through a paramagnetic beads method (Zhijiang Biotechnology Co., Ltd., Shanghai, China). The concentration and purity (A260/A280 ratio) of nucleic acid for each RNA extract were evaluated with a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, USA). A260/A280 ratio of pure RNA products ranged from 1.8 to 2.0 (8). Extracted RNA.....
Document: RNA was extracted simultaneously everyday by using the Automated Nucleic Acid Extraction System through a paramagnetic beads method (Zhijiang Biotechnology Co., Ltd., Shanghai, China). The concentration and purity (A260/A280 ratio) of nucleic acid for each RNA extract were evaluated with a Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, USA). A260/A280 ratio of pure RNA products ranged from 1.8 to 2.0 (8). Extracted RNA was tested by rRT-PCR using IAV Detection kit (Zhijiang Biotechnology Co., Ltd.) with ABI 7500 Real-Time PCR instrument (Foster City, CA, USA). Threshold cycle (Ct) values being negatively related to concentration of nucleic acid were recorded. Operation and result assessment were conducted in accordance with the manufacturers' instructions. Qualified negative, positive, and internal controls were the premise of results validity.
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