CRISPR-Cas13 a-based bovine viral diarrhea virus detection method
A technology of bovine viral diarrhea and virus, which is applied in the field of detection of bovine viral diarrhea virus, can solve the problems of high cost, difficult to deal with on-site and large-scale detection, and high probability of false positive, achieving high application value and high accuracy , the effect of rapid detection
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[0027] The present invention will be further described in detail below in conjunction with specific embodiments, and the given examples are only for clarifying the present invention, not for limiting the scope of the present invention.
[0028] The inventor expresses and purifies LwCas13a protein with LwCas13a as the research center, and at the same time, designs a specific crRNA according to the 5'UTR conserved region of the reported BVDV gene sequence. The RNA obtained from BVDV was used to verify that Cas13a also has RNA-cleaving activity in vitro, and can specifically and accurately detect BVDV, and the invention was completed on this basis.
[0029] Step 1: Primer design for the detection method of bovine viral diarrhea virus based on CRISPR-Cas13a
[0030] 1. LwCas13a gene primer design
[0031] The full-length gene sequence of pC013-Twinstrep-SUMO-LwCas13a was obtained on addgene, and the full-length primers of LwCas13a protein were designed with Premier5 software. Th...
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