Method for obtaining target nucleic acid form mixed nucleic acid
A nucleic acid and target technology, applied in the identification of low-abundance bacterial species in human metagenome samples, can solve the problems of high sequencing cost, difficult analysis of low-abundance microorganisms, and large amount of data, so as to improve the identification rate and reduce sequencing data. wasteful effect
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[0022] (1) Material preparation
[0023] A small amount of meta samples of human origin were obtained, and the meta samples obtained were human skin meta samples containing a total of 32 ng of nucleic acids at a concentration of 1.1 ng / μl.
[0024] (2) The method can be divided into the following steps
[0025] 1. Enrich a trace amount of meta nucleic acid
[0026] This step removes more than 90% of the human-derived DNA, greatly increasing the content of meta DNA, and realizing the so-called enrichment of a small amount of meta DNA.
[0027] Specifically, MBD2-Fc was bound to magnetic beads coated with protein A, combined with a vertical suspension apparatus at room temperature for 10 min, washed twice with binding buffer, the binding buffer was composed of 10 mM Tris, pH7.5, 1 mM EDTA, Composition of 0.2% Tween20 and 300mM NaCl, then resuspend MBD2-Fc-magnetic beads with binding buffer, combine with DNA sample for 50min, place the reaction tube on the magnetic stand until ...
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