2.7. Statistical analysisRaw fastq files were demultiplexed and quality-filtered by usingQIIME (version 1.17) with the following criteria: (1) the 250 bpreads were truncated at any site to receive an average quality score<20 over a 10 bp sliding window, with discarding the truncatedreads that were shorter than 50 bp; (2) as exacting barcodematching and two nucleotide mismatch in primer matching, readsthat containing ambiguous characters were removed; (3) only thesequences that overlap longer than 10 bp were assembled accordingto their overlap sequence, and reads which could not beF ¼ 2△△Ct, △△Ct ¼ (Ct, target gene e Ct, reference gene) e (Ct, targetgene e Ct, reference gene)control [35]. Differences between experimentalgroup and control were analyzed using SPSS (version 18.0) statisticalsoftware (SPSS Inc., Chicago, IL, USA) using post-hoc test,Dunnett's. A p-value of less than 0.05 or 0.01 was statisticallyconsidered significantly.
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