The invention relates to a method for constructing a bacterium 16S rDNA overall-length high-
throughput sequencing
library, and relates to the field of
microorganism high-
throughput sequencing. The method comprises the steps of firstly, adding unique molecular identifiers (UMIs) which are composed of random base groups to the two ends of each 16S rDNA template molecule, conducting PCR amplificationto obtain multiple copied 16S rDNA overall-length amplicons with two ends containing specific UMIs, randomly breaking and connecting joints of the sequencing
library, and conducting paired-end sequencing to obtain the UMI sequences and corresponding 16S rDNA fragment sequences; besides, through cyclizing of the 16S rDNA overall-length amplicons and sequencing, obtaining UMI
pairing information oftwo ends of the same molecule; conducting reads
assembly according to the same UMI and
pairing UMI information to increase the splicing length accordingly, and finally conducting
assembly to obtain 16S rDNA overall-length sequence information. The method is low in cost, high in accuracy and suitable for high-
throughput sequencing platforms; meanwhile, the method can effectively eliminate the influence of PCR amplification preferences, amplification errors and sequencing errors through the UMIs, and thus the bacterium
population abundance in a sample is more precisely quantified.