Primer for detecting botryosphaeria dothidea of kiwi fruit and detection method and application
A technology of kiwi fruit and pathogenic bacteria, applied in the field of high-sensitivity specific primers and detection, can solve the problems of unsatisfactory sensitivity, etc., and achieve the effect of easy implementation and simple detection process
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Examples
Embodiment 1
[0046] (1) DNA is extracted from kiwi fruit; the specific method is:
[0047] 1) Put an appropriate amount of kiwi fruit into a 1.5ml EP tube, add liquid nitrogen, and quickly grind it into powder with a grinding rod;
[0048] 2) Add 550 μL of CTAB extract solution preheated at 65°C, shake vigorously, place in a 65°C incubator for 1 hour, shake and mix once every 10 minutes
[0049] 3) Add 550 μL of chloroform-isoamyl alcohol (24:1 v / v), shake vigorously and mix well, let stand for 10 min, and centrifuge at 13000 r / min for 5 min;
[0050] 4) Take the supernatant, add 600 μL ice ethanol, centrifuge at 13000 r / min for 5 min, pour off the supernatant, and wash the precipitate with 75% ethanol; repeat this step at least once;
[0051] 5) Empty centrifuge at 7500r / min for 3min, suck off excess liquid at the bottom with a pipette, and let it dry naturally at room temperature;
[0052] 6) After drying, add an appropriate amount of sterilized ddH 2 O was fully dissolved, and then s...
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Description
Claims
Application Information
- IPC
- C12Q1/6895; C12Q1/686; C12Q1/04; C12N15/11; C12R1/645
- CPC
- C12Q1/686; C12Q1/6895; C12Q2565/125
- Inventors
- 胡容平; 石军