Primer for RPA rapid test of salmonella carrying sulfonamides resistance gene and application of primer
A Salmonella and drug-resistant gene technology, applied in the field of RPA rapid detection primers, can solve the problems of unsatisfactory operation procedures and detection time specificity detection, and achieve good intra-species conservation and good inter-species specificity
Image
Examples
Embodiment 1
[0028] Step (1). Sample DNA extraction
[0029] Take 1ml of the cultured bacteria solution and add it to a 1.5ml centrifuge tube, centrifuge at 12000g for 1min, discard the supernatant, add 400μl lysate to the centrifuge tube, mix well, add 600μl phenol-chloroform-isoamyl alcohol mixed solvent, shake vigorously, and centrifuge at 12000g 10min; take the supernatant, add chloroform-isoamyl alcohol mixed solvent equal to the volume of the supernatant, shake vigorously, and centrifuge at 12000g for 10min; take the supernatant, add chloroform equal to the volume of the supernatant, shake vigorously, Centrifuge at 12,000 g for 10 min; take the supernatant, add isopropanol 0.8 times the volume of the supernatant, and centrifuge at 12,000 g for 10 min; take the precipitate, wash it once with ethanol with a volume content of 70%, and then dry it at room temperature or 50°C Dry for 20 minutes, then add 80 μl of TE buffer solution with a pH value of 8.0 to dissolve to obtain a DNA extrac...
Embodiment 2
[0042] Step (1). Sample DNA extraction
[0043] Take 1ml of the cultured bacteria solution and add it to a 1.5ml centrifuge tube, centrifuge at 12000g for 1min, discard the supernatant, add 400μl lysate to the centrifuge tube, mix well, add 600μl phenol-chloroform-isoamyl alcohol mixed solvent, shake vigorously, and centrifuge at 12000g 10min; take the supernatant, add chloroform-isoamyl alcohol mixed solvent equal to the volume of the supernatant, shake vigorously, and centrifuge at 12000g for 10min; take the supernatant, add chloroform equal to the volume of the supernatant, shake vigorously, Centrifuge at 12,000 g for 10 min; take the supernatant, add isopropanol 0.8 times the volume of the supernatant, and centrifuge at 12,000 g for 10 min; take the precipitate, wash it once with ethanol with a volume content of 70%, and then dry it at room temperature or 50°C Dry for 20 minutes, then add 80 μl of TE buffer solution with a pH value of 8.0 to dissolve to obtain a DNA extrac...
Embodiment 3
[0056] Step (1). Sample DNA extraction
[0057] Take 1ml of the cultured bacteria solution and add it to a 1.5ml centrifuge tube, centrifuge at 12000g for 1min, discard the supernatant, add 400μl lysate to the centrifuge tube, mix well, add 600μl phenol-chloroform-isoamyl alcohol mixed solvent, shake vigorously, and centrifuge at 12000g 10min; take the supernatant, add chloroform-isoamyl alcohol mixed solvent equal to the volume of the supernatant, shake vigorously, and centrifuge at 12000g for 10min; take the supernatant, add chloroform equal to the volume of the supernatant, shake vigorously, Centrifuge at 12,000 g for 10 min; take the supernatant, add isopropanol 0.8 times the volume of the supernatant, and centrifuge at 12,000 g for 10 min; take the precipitate, wash it once with ethanol with a volume content of 70%, and then dry it at room temperature or 50°C Dry for 20 minutes, then add 80 μl of TE buffer solution with a pH value of 8.0 to dissolve to obtain a DNA extrac...
PUM
Login to View More Abstract
Description
Claims
Application Information
- IPC
- C12Q1/68; C12Q1/10; C12N15/11; C12R1/42
- CPC
- C12Q1/6844; C12Q1/689; C12Q2600/106; C12Q2521/507; C12Q2521/101; C12Q2565/125; Y02A50/30
- Inventors
- 曲道峰; 韩剑众