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Reagent kit for Bcl-w gene hybridization in situ, detection method and uses thereof

A technology of in situ hybridization and detection method, which is applied in the direction of biochemical equipment and methods, microbe determination/inspection, etc., and can solve the problems that detection technology has not been reported.

Inactive Publication Date: 2009-05-13
NATUREGEN BIOTECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But, there is no report about the in situ hybridization detection kit and detection technology of Bcl-w gene

Method used

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  • Reagent kit for Bcl-w gene hybridization in situ, detection method and uses thereof
  • Reagent kit for Bcl-w gene hybridization in situ, detection method and uses thereof
  • Reagent kit for Bcl-w gene hybridization in situ, detection method and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Prepare the in situ hybridization kit of this embodiment according to conventional methods, the kit includes hybridization probes, markers, and instructions designed with the BCL-W gene as the detection target gene, wherein:

[0039] Digoxigenin was selected as the probe label in this embodiment.

[0040] Kit composition:

[0041]Digestive solution 100μL / tube 1 tube / box Colorless transparent liquid

[0042] Protective solution 100μL / tube 1 tube / box Colorless transparent liquid

[0043] Pre-hybridization solution 1300μL / tube 2 tubes / box Colorless transparent liquid

[0044] Sense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0045] Antisense hybridization solution 10μL / tube 1 tube / box Colorless transparent liquid

[0046] Blocking solution 1000μL / tube 1 tube / box Colorless transparent liquid

[0047] Alkaline phosphatase antibody 1μL / tube 1 tube / box Colorless transparent liquid

[0048] Chromogen A 175μL / tube 1 tube / box Yellow liquid

...

Embodiment 2

[0090] Specimen processing:

[0091] 1). Use a 10ml centrifuge tube to fill 4.5ml of lymphocyte separation solution, then slowly add 3ml of anticoagulated blood into the centrifuge tube containing lymphocyte separation solution (blood: lymphocyte separation solution = 1:1.5), 2000r / min Centrifuge for 10min

[0092] 2). Draw the white blood cells in the middle layer into another centrifuge tube, then add about twice the amount of 1× buffer I to this tube, mix well, and centrifuge at 1500g / min for 10min

[0093] 3). Discard the supernatant. Add about twice the 1× buffer I to the pellet, mix well, and centrifuge at 1500g / min for 10min

[0094] 4). Discard the supernatant, and absorb the excess liquid from the mouth of the test tube with paper towels. Then the precipitate was made into a suspension, dropped on a glass slide, and allowed to dry naturally. (Hospitals with conditions can use the film-making machine to make films.) With 3ml of blood, 4 films can be made.

[0095] ...

Embodiment 3

[0098] Prepare the reagents in the kit to the concentration used

[0099] 1). Dilute 10× buffer I with triple distilled water at 1:10 to 1× buffer I;

[0100] 2). Dilute 20× buffer II with triple distilled water at 1:10 to 2× buffer II;

[0101] Dilute 1:100 into 0.2×buffer II; dilute 1:200 into 0.1×buffer II;

[0102] 3). Dilute 10× buffer III with triple distilled water at 1:10 to 1× buffer III;

[0103] 4). Dilute 10× buffer IV with triple distilled water at a ratio of 1:10 to form × buffer IV (take 10 mL each of 1#, 2#, and 3#, and add water to 100 mL).

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Abstract

The invention discloses an in situ hybridization detection kit for early metastasis of peptic cancer and other cancer types, which comprises hybrid probes and labels. The invention also discloses a method for performing in situ hybridization detection on Bcl-w genes related to peptic cancer generation and metastasis by utilizing the kit. The method comprises the following steps: (1) RNA to be detected in a substrate contacts the hybrid probes to form a hybrid complex under the condition that the hybrid probes and target sequences can form a stable hybrid complex; and (2) the hybrid complex is detected. The kit and the detection method can detect the expression amount of the Bcl-w genes in the genetic level, is earlier than detection by the prior image medical means, and can realize the real early diagnosis and metastasis and relapse forecast of peptic cancer and other cancer types. Meanwhile, the detection method is simple and convenient, and is convenient to popularize and apply.

Description

technical field [0001] The invention relates to the field of biological detection and disease diagnosis, more specifically, it relates to the detection technology of genes related to occurrence and metastasis of digestive tract and other cancers. Background technique [0002] According to the survey by the national authoritative organization, there are about 6 million cancer patients in my country, 1.6 million new cancer patients and 1.6 million deaths every year. There are nearly 84 million cancer patients in the world, and 8 million people die of cancer every year. This number will double in 2020, which is a set of terrifying figures. Scientists and tumor clinicians have made unremitting efforts in the diagnosis and treatment of cancer, but the mortality rate of cancer is very high, mainly due to the metastasis of cancer. On October 5 last year, the National Institutes of Health (NIH), the Cancer Institute (NCI) and several other units jointly completed the "Annual Report...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 裘建英张云福
Owner NATUREGEN BIOTECH SHANGHAI