The DNA sequence of this marker gene is compared to a reference database for the identification of bacteria based on the present or absent the sequence in the database. The gold standard for microbe identification is an advanced technique for bacterial identification, is to identifying specific marker genes in their chromosome-a fingerprint gene. Bacteria can now be identified to species level for the first time. This landscape has been significantly changed by novel DNA-based technologies without growing pure culture. It is true that determining the functions of bacteria is difficult, however there are several approaches for doing so. The majority of the species discovered are new and unknown, and many of them belong to unnamed genera. This will be extremely crucial in the coming years, given the global trend toward advanced water resource recovery systems for sustainable development. To make appropriate decisions about the management of the treatment systems, detail information about the microorganism is essential. It is only possible to determine whether the correct processes are in right place when the identity and function of the majority of microorganisms are known, which is difficult to do today. Microbial community surveillance might be used as a standard approach of controlling and optimising wastewater treatment processes. Diverse microorganisms are in the systems however, only a few of them have information about their identification and functions that are involved in the treatment system. Each of these bacteria contributes to the treatment process in a different way, ensuring that there is little to no influence on the environment. There is a diversity of microorganisms employed in sewage treatment, however, the exact number is unknown, which play a vital role in wastewater treatment. Microorganisms are a crucial component of wastewater treatment systems, as they remove pollutants and disease-causing pathogens, recycle nutrients and energy, and produce clean water. If the effluent is not properly treated, it may cause serious harm to humans and the environment. Industrial waste, chemical effluents, untreated sewage, human waste, animal waste, garbage, and other pollutants are the most common sources of contamination of water. Waterborne infectious diseases are caused by viruses and bacteria that are ingested into the body through contaminated water or contact with faeces. The robustness of NS-watermark barcodes, together with their scalable design and compatibility with low-cost massive synthesis, makes them promising for present and future sequencing applications requiring massive labeling, such as long-read single-cell RNA-Seq. This falls in the range of the index hopping rate of established, high-accuracy Illumina sequencing, despite the increased number of tags and the relatively low accuracy of both microarray-based synthesis and long-read sequencing. We recovered the identity of more than 86% of the barcodes, with a crosstalk rate of 0.17% (i.e., one misassignment every 584 reads). To our knowledge, this is the largest number of distinct, non-random tags ever sequenced in parallel and the first report of microarray-based synthesis as a source for large oligonucleotide pools for barcoding. Here, we apply so-called NS-watermark barcodes, whose error correction capability was previously validated in silico, in a proof of concept where we synthesize 3840 NS-watermark barcodes and use them to asymmetrically tag and simultaneously sequence amplicons from two evolutionarily distant species (namely Bordetella pertussis and Drosophila mojavensis ) on the ONT MinION platform. Nucleic-acid barcoding is an enabling technique for many applications, but its use remains limited in emerging long-read sequencing technologies with intrinsically low raw accuracy.
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