title: bacteria
design: 1) Collect and purify genomic DNA;2) Covaris was used to fragment genomic DNA and construct genomic sequencing library;3) Connect A & B connectors;4) Screening and removing joint self-contiguous segments;5) Fragments were screened by agarose gel electrophoresis, and fragments with one end being A joint and one end being B joint were retained;6) Sodium hydroxide denatures, producing single-stranded DNA fragments.Bridge PCR7) One end of the DNA fragment is complementary to the primer base and fixed on the chip;8) The other end randomly complements another nearby primer and is also fixed to form a "bridge";9) PCR amplification to produce DNA clusters;10) DNA amplicon linearization into a single strand.11) Add modified DNA polymerase and dNTP with 4 kinds of fluorescent labels, and only one kind of base is incorporated in each cycle;12) The surface of the reaction plate is scanned by laser to read the types of nucleotides polymerized in the first reaction of each template sequence;13) The "fluorophores" and "terminating groups" are chemically cut to restore the 3' end viscosity and continue to polymerize the second nucleotide;14) Statistical results of fluorescence signals collected in each round to obtain the sequence of template DNA fragments.