RNA-Seq of anaerobic sludge
1) Firstly, sequence end repair is performed on the above amplification product by removing the prominent base at the 5' end of the DNA sequence and adding a phosphate group to complete the missing base at the 3' end by End Repair Mix 2 in the kit.2) Adding an A base to the 3' end of the DNA sequence to prevent self-association of the DNA fragment and to ensure that the target sequence can be attached to the sequencing junction (the sequencing junction has a prominent T base at the 3' end).3) Addition of a sequencing junction containing a library-specific tag (i.e. Index sequence) at the 5' end of the sequence to enable the DNA molecule to be immobilized on the Flow Cell.4) purify the library system after the addition of the junction by removing the self-associated fragment by magnetic bead screening using BECKMAN AMPure XP Beads.5) PCR amplification of the above junction-linked DNA fragments to enrich the sequencing library template, and re-purification of the library enrichment product using BECKMAN AMPure XP Beads.6) Final selection and purification of the library by 2% agarose gel electrophoresis.Translated with www.DeepL.com/Translator (free version)