JCNRJournal of Clinical and Nursing Research2208-36852208-3693Bio-Byword Scientific Publishing Pty. Ltd.10.26689/jcnr.v9i10.12670ArticleStudy on a Rapid Extraction and Detection Method for 16S rRNA of Intestinal Florahttps://artdesignp.com/journal/JCNR/9/10/10.26689/jcnr.v9i10.12670WangYuanyuan20259102025-11-10This study established a rapid extraction method for the 16S rRNA gene of intestinal flora. Combined with next-generation sequencing (NGS) technology, this method can be applied to intestinal microbial ecology analysis. The fecal sample was mixed with lysis buffer, incubated at 90°C for 15 minutes, vortexed, and then centrifuged. The supernatant was collected, and specific primers were added for PCR amplification. The PCR products were purified, and index tags were established for the targets to obtain libraries. After library purification and quality inspection, the libraries could be loaded for sequencing. Under optimized conditions: when the number of PCR cycles was 25 and the initial amount of DNA for library construction was 12.5 ng, fewer chimeras were generated. The library size was 500–700 bp, and there were no primer dimers of approximately 120 bp. This method is rapid, accurate, and sensitive, and can be used for the analysis and detection of the 16S rRNA gene of intestinal flora.

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