Bovine babesiosisbigemina nested PCR specific primer as well as detection kit and nested PCR detection method
A technology of Babesia double buds and a detection kit, which is applied in the field of biology, can solve the problems of high false positive rate, achieve high sensitivity, improve specificity and sensitivity, and detect conveniently and quickly
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[0038] Set up a nested PCR detection kit that includes:
[0039] (1) EP dof tube: The reaction tube contains 10×PCR Buffer, deoxyribonucleoside triphosphate (dNTP), magnesium chloride (Mgcl 2 ).
[0040] (2) Primer F1, primer R1, primer F2, primer R2. Primer F1, primer R1, primer F2, and primer R2 are DNA fragments synthesized by a DNA synthesizer according to the following base sequences:
[0041] Primer BF1: 5'-CAT-CTA-ATT-TCT-CTC-CAT-ACC-CCT-CC-3';
[0042] Primer BR1: 5'-CCT-CGG-CTT-CAA-CTC-TGA-TGG-CAA-AG-3';
[0043] Primer BF2: 5'-CGC-AAG-CCC-AGC-AGG-CCC-CGG-TGC-3';
[0044] Primer BR2: 5'-CCG-ACC-TGG-ATA-GGC-TGT-GTC-ATG-3'.
[0045] (3) Positive control: This control is a recombinant plasmid containing the target gene fragment, which is constructed by our laboratory. The method is as follows: clone the amplified product into pUCm-T to transform DH5-α competent cells, obtain a positive recombinant plasmid after enzyme digestion identification and sequencing, and the ...
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