In-situ hybridization detection kit for MICRORNA (MICRO Ribonucleic Acid)-92A level at pathologic evolution early stage of colon cancer as well as microRNA-92A in-situ hybridization detection method and application of microRNA-92a to preparation of in-situ hybridization detection kit for colon cancer
A detection kit and in situ hybridization technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of non-decreasing mortality, drug resistance of tumor cells, failure of anti-cancer battle, etc., and achieve specificity Strong, easy to operate, high sensitivity effect
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Embodiment 1
[0052] The in situ hybridization kit of this example was prepared according to conventional methods, and the kit included hybridization probes designed with MICRORNA-92A, markers, and instructions, wherein the probe marker of this example was digoxin.
[0053] Kit hybridization solution composition:
[0054] 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 liquid Chro...
Embodiment 2
[0062] The implementation process of applying nucleic acid in situ hybridization detection method to the expression level of MICRORNA-92A in each group of blood samples:
[0063] 1).Take two specimens to be tested;
[0064] 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;
[0065]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;
[0066] 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 box tightly, and...
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