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Full-length 16S rRNA gene sequencing try carried out playing with an improved form of all of our method for man-made long-understand sequencing19. To switch amplification away from bacterial 16S rRNA gene diversity to the long-realize sequencing, i replaced the newest 1492r primer in past times used18 on the 1391r primer48 as it has greatest visibility of your known microbial diversity. Simultaneously, i replaced the brand new Taq polymerase used in first amplification of your book molecular identifier (UMI) tagged themes having a great proofreading polymerase (Second Fig. 16). So it increased the fresh percentage of error-free reads almost five-bend, and therefore greatly enhances our capacity to resolve full-length 16S rRNA ASVs (FL-ASVs) out of reduced abundant taxa as the at the least two similar sequences try necessary to take care of an Fl-ASV18.
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