Horizontal in-situ hybridization detection kit and detection method as well as application for MICRORNA-143 in earlier stage of cancer pathologic evolution
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 the advantages of convenient operation, strong specificity and high sensitivity Effect
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Embodiment 1
[0053] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes designed with MICRORNA-143, markers, instructions, wherein:
[0054] Digoxigenin was selected as the probe label in this embodiment.
[0055] Kit hybridization solution composition:
[0056] 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 yell...
Embodiment 2
[0064] The implementation process of applying nucleic acid in situ hybridization detection method to the expression level of MICRORNA-143 in each group of blood samples:
[0065] 1).Take two specimens to be tested;
[0066] 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;
[0067] 3). Wash with 0.2% protection solution (protection solution 1ml plus 1× buffer Ⅰ, 99ml is the used concentration) for 10 minutes, three-distilled water for 5 minutes (the above process is carried out in a glass tank), take out the slide and let it naturally dry;
[0068] 4). Put the slides into the humidity box, add 25 μL / slice of pre-hybridization solution (add to the place where there are cells), cover with a cover glass, cover the humidity box tightly, and pu...
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