MRNA-level in-situ hybridization detection kit for KLOTHO gene in pancreas precancerous period of KLOTHO gene in prophase of pancreas cancer detecting method and application
A detection kit and in situ hybridization technology, which is applied in the related detection technology field of mRNA expression changes, can solve problems such as non-decreasing mortality, drug resistance of tumor cells, failure of anti-cancer battle, etc., and achieves convenient operation and strong specificity , the effect of high sensitivity
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Embodiment 1
[0054] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes, markers, and instructions designed with the KLOTHO gene as the detection target gene, wherein:
[0055] Digoxigenin was selected as the probe label in this embodiment.
[0056] Kit hybridization solution composition:
[0057] 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 Chromoge...
Embodiment 2
[0065] The implementation process of applying the nucleic acid in situ hybridization detection method to the KLOTHO gene expression of each group of blood samples:
[0066] 1).Take two specimens to be tested;
[0067] 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;
[0068] 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;
[0069] 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 t...
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