Human DNA hijacking microbiota surveys: causes and consequences in colon related 16s rRNA amplicon sequencing.

ABSTRACT: The efficiency of polymerase chain reaction (PCR) decreases under suboptimal conditions, such as low template concentration combined with high concentrations of similar sequences. Under these circumstances, mis-priming can occur, leading to the generation of erroneous copies. Specifically, in 16S amplicon sequencing of human intestinal biopsy samples, host off-target sequences are frequently generated and subsequently sequenced, particularly when the commonly used V3-V4 primers are employed. This issue not only introduces errors in data interpretation but also results in the unnecessary consumption of sequencing depth. In response to this challenge, we analysed over 1,300 publicly available V3-V4 amplicon sequences related to the human colon, profiling the colon microbiota while elucidatin

SUBMITTER: Di Gloria L 

PROVIDER: S-EPMC12455517 | biostudies-literature | 2025

REPOSITORIES: biostudies-literature