Cancer pathologic evolution early-stage Gankyrin gene mRNA level in-situ hybridization detection kit, detection method and applications thereof
A detection kit and in situ hybridization technology, applied in the field of biological detection, can solve problems such as human and environmental damage, and achieve the effects of convenient operation, strong specificity and high sensitivity
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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 Gankyrin gene mRNA, markers, instructions, wherein:
[0053] Digoxigenin was selected as the probe label in this embodiment.
[0054] Kit hybridization solution composition:
[0055] Digestive solution 100μL / tube 1 tube / box Colorless transparent liquid
[0056] Protective solution 100μL / tube 1 tube / box Colorless transparent liquid
[0057] Pre-hybridization solution 1300μL / tube 2 tubes / box Colorless transparent liquid
[0058] Sense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid
[0059] Antisense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid
[0060] Blocking solution 1000μL / tube 1 tube / box Colorless transparent liquid
[0061] Alkaline phosphatase antibody 1μL / tube 1 tube / box Colorless transparent liquid
[0062] Chromogen A 175μL / tube 1 tube / box Yellow liquid
[0063...
Embodiment 2
[0077] The implementation process of applying the nucleic acid in situ hybridization detection method to the Gankyrin gene mRNA expression of each group of blood samples:
[0078] 1).Take two specimens to be tested;
[0079] 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;
[0080] 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;
[0081] 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 humidifyin...
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