LET-7micro ribose nucleic acid (MIRNA) level in-situ hybridization detection kit for pathologic evolution early stage of various cancer, and detection method and application
A detection kit and in situ hybridization technology, applied in the field of related detection technology, can solve the problems of non-decreasing mortality, drug resistance of tumor cells, failure of anti-cancer war, etc., and achieve convenient operation, high sensitivity and strong specificity Effect
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Embodiment 1
[0052] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes designed with LET-7MIRNA, markers, instructions, wherein:
[0053] Digoxigenin was selected as the probe label in this embodiment.
[0054] Kit hybridization solution composition:
[0055] digestive juice 100μL / tube 1 tube / box colorless transparent liquid protective fluid 100μL / tube 1 tube / box colorless transparent liquid Pre-hybridization solution 1300μL / tube 2 tubes / box colorless transparent liquid Right-sense hybridization solution 10μL / tube 1 tube / box colorless transparent liquid antisense hybridization solution 10μL / tube 1 tube / box colorless transparent liquid blocking solution 1000μL / tube 1 tube / box colorless transparent liquid Alkaline phosphatase antibody 1 μL / tube 1 tube / box colorless transparent liquid Chromogen A 175μL / tube 1 tube / box yellow...
Embodiment 2
[0063] The implementation process of applying the nucleic acid in situ hybridization detection method to the LET-7MIRNA expression level of each group of blood samples:
[0064] 1).Take two specimens to be tested;
[0065] 2). Add 50 ml of digestive solution (100 μL of digestive solution plus 99.9 ml of 1× buffer Ⅰ, which is the concentration used) in a glass tank, preheat in a water bath at 37°C for 10 minutes, put 16 slides in, and treat at 37°C for 12 minutes , and then washed with 1× buffer I for 5 min;
[0066] 3). Use 0.2% protection solution (protection solution 1ml plus 1× buffer , 99ml is the concentration used), washed for 10 minutes, washed with three-distilled water for 5 minutes (the above process was carried out in a glass tank), took out the slide, and let it dry naturally;
[0067] 4). Put the slides into a humidifying box, add 25 μL / slice of pre-hybridization solution (add to the place where there are cells), cover with a cover glass, cover the humidifying ...
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