Pseudomonas plecoglossicida aptamers and screening method therefor
A Pseudomonas mutans, screening method technology, applied in biochemical equipment and methods, DNA preparation, DNA / RNA fragments, etc., can solve problems such as inability to fully explain pathogenicity
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[0036] The present invention takes Pseudomonas mutans as an object, adopts reverse screening and other SELEX technologies to screen nucleic acid aptamers that can specifically recognize Pseudomonas mutans, and then verifies the affinity of the nucleic acid aptamers.
[0037] 1 SELEX screening of nucleic acid aptamers
[0038] (1) Bacteria treatment: take the cultured Pseudomonas mutans centrifuge tube, centrifuge at 6000r / min for 5min, discard the supernatant, wash the bacteria 3 times, add sterile culture medium and mix evenly, measure the OD value of the bacteria solution, Then take the bacteria content of about 4 × 10 8 Centrifuge the Pseudomonas mutans solution at 6000r / min for 5min, discard the supernatant, wash the bacterial pellet three times with 0.9% normal saline, wash the bacterial pellet once with 1×binding buffer, and finally add 100μL of 2×binding buffer Resuspended.
[0039] (2) Binding: take ssDNA random library, dilute to 2μmol / L100μL with 2×binding buffer, ...
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