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Use of human Down's syndrome DNA sequence in detection on the number of human cells implanted in mouse

A technology for Down syndrome and DNA sequence, applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial determination/inspection, etc., can solve the problems of low specificity, complex methods, poor repeatability, etc.

Active Publication Date: 2015-04-08
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although there have been methods for detecting human cells in tissues based on PCR technology in the past, there are defects such as complex methods, poor reproducibility, and low specificity.

Method used

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  • Use of human Down's syndrome DNA sequence in detection on the number of human cells implanted in mouse
  • Use of human Down's syndrome DNA sequence in detection on the number of human cells implanted in mouse
  • Use of human Down's syndrome DNA sequence in detection on the number of human cells implanted in mouse

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1. Obtaining of a kit for detecting human cells implanted in mice

[0023] 1. Sequence-specific analysis of Down syndrome

[0024] By analyzing the NCBI (National Center for Biotechnology Information) GenBank database to screen human-specific genome sequences, the human-specific Down syndrome sequence (Human Down Syndrome region of chromosome21, clone A37A2-1F8) on the 21st pair of chromosomes was finally selected. .ACCESSION: U50569, whose sequence is shown in sequence 3 in the sequence table), download the sequence, and use the sequence to perform Blast on rat and mouse genome sequences. The mouse genome sequence has great specificity and is an ideal target sequence to detect the number of human cells implanted in the mouse.

[0025] 2. Design of primer pair sequences

[0026] Based on the Down syndrome sequence as a template, multiple pairs of real-time PCR primers were designed with Generunner software, and these primer sequences were used to perform Blast...

Embodiment 2

[0033] Example 2. The kit for detecting human cells implanted in mice The effect of detecting the distribution of human mesenchymal stem cells in mice with myocardial infarction after intravenous injection was verified. .

[0034] The specific method of preparing the acute myocardial infarction model in mice (BALB / c was purchased from the Animal Center of Southern Medical University) is as follows

[0035] 1. Experimental animals

[0036] 11-12 weeks balb / c mice, body weight 25-30g

[0037] 2. Surgical instruments

[0038] 2% sodium pentobarbital solution; dermatome; adhesive tape and pins for fixation; iodine, alcohol; surgical blades; Needle, curved forceps, straight forceps, non-invasive suture needles, operating table, 8 / 0 sutures, 10 bundles of 5 / 0 nylon threads; small animal ventilator, electrocardiogram machine; incandescent lamp 60w.

[0039] 3. Anesthesia, immobilization, monitoring

[0040]Inject 2% pentobarbital sodium solution (50mg / kg) intraperitoneally, obse...

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Abstract

The invention discloses a method for detecting the number of human cells implanted in a mouse and a kit utilizing the method. The kit utilizing the method for detecting the number of human cells implanted in a mouse comprises a primer pair for amplifying a human Down's syndrome DNA sequence fragment. The human Down's syndrome DNA sequence fragment has the sequence 3 in the sequence table. The method and the kit realize specific detection on the number of human cells in mouse tissue. Through use of the primers having high specificity, the kit has the advantages of high specificity, strong sensibility and good repeatability.

Description

technical field [0001] The present invention relates to the application of human Down's syndrome DNA sequence as a target sequence for detecting the number of human cells implanted in a mouse, and the method for detecting the number of human cells implanted in a mouse with a human Down's syndrome DNA sequence and based on this Application of the method kit. Background technique [0002] Mice (including rats and mice) are commonly used biomedical experimental animals, and are widely used in experimental research on human cancer cell metastasis and human stem cell transplantation therapy. In the experiment, a certain number of human cells need to be injected into the mouse body. Accurately knowing the distribution of these cells in the mouse body is very important for studying the characteristics of the cells and judging the effect of drug treatment, but there has been a long-term lack of a sensitive and stable method to determine these human cells Distribution of cancer cell...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888
Inventor 吴耀炯谢振华宋鹏跃
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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