Multiplex amplification system and detection kit for short tandem repeat sequence of mouse

A composite amplification system and short tandem repeat technology, applied in the biological field, can solve the problems of stutter peak height, inability to judge, and unfavorable cross-contamination between mouse cells.

Active Publication Date: 2016-06-08
NAT INST FOR FOOD & DRUG CONTROL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the STR sites used in this patent are all 2nt. repeats, resulting in a stutter peak height, which is not conducive to ju

Method used

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  • Multiplex amplification system and detection kit for short tandem repeat sequence of mouse
  • Multiplex amplification system and detection kit for short tandem repeat sequence of mouse
  • Multiplex amplification system and detection kit for short tandem repeat sequence of mouse

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Experimental program
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Effect test

Embodiment 1

[0123] Below is that we adopt the present invention to mix 19 routine mouse cells, 1 routine rat cell, 1 routine hamster cell, 1 routine woodchuck cell, 1 routine guinea pig cell, 1 routine human source DNA and 1 routine by the ratio of 5:1 And the specific implementation of the detection of mouse and human cell samples.

[0124] 1. DNA extraction

[0125] Chelex-100 and DNA extraction kit (Tiangen Biochemical) were used for DNA extraction. The operation steps were in accordance with the instructions. After the DNA extraction was completed, the DNA was quantified with a UV spectrophotometer and diluted to 2 ng / μl.

[0126] 2.PCR

[0127] 2.1 Reaction system:

[0128] A total of 12 pairs of primers for 9 mouse STR sites, sex determination site Jarid1 and 2 human STR sites were dissolved respectively and made into a working solution with a concentration of 10 μM, and then prepared into a primer mixture according to the volume ratio in Table 2 ( Primermix): See the sequence li...

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Abstract

The invention relates to a multiplex amplification system and a detection kit for a short tandem repeat sequence of a mouse, and belongs to the technical field of biological detection. According to the multiplex amplification system and the detection kit for the short tandem repeat sequence of the mouse, nine STR (Short Tandem Repeat) loci, which have high sensitivity and amplification specificity, of the mouse are selected; the STR loci are amplified once; the loci amplified by the multiplex amplification system are some STR loci which can be used for identifying a cell line of the mouse; when the cell line of the mouse is identified, the multiplex amplification system and the detection kit have the advantages of being quick, being simple to operate and accurate in result, being convenient for large-scale popularization, and the like. By using a method, along with the successful development of the kit, the batch production can be realized; a working condition can be modeled; automatic equipment exists in all processes; human operation experience is not depended on any more; the automation can be realized; a required time is short; a whole detection time needs approximate 6 hours.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a compound amplification system for identifying multiple short tandem repeat sequences in mice and a kit thereof, which can rapidly and accurately perform STR typing of mouse cells and further identify mouse cell line types. Background technique [0002] In 2009, an editorial in Nature pointed out: Among the thousands of biological laboratories using cell lines, many laboratories do not know that between 1 / 5-1 / 3 of the commonly used cell lines may have been It's not what it used to be. Numerous studies over the past 25 years, combined with experience with cell banks in the US, UK, Germany, and Japan have found that 18-36% of cultures contain misidentified species or other cell types. [0003] In May 2010, the ATCCSDO (Standards Development Organization) working group published a forward-looking report entitled "Cellline misidentification: the beginning of the end" in the journal Natur...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 孟淑芳樊金萍徐苗吴雪伶吕悦心陈初光赵翔冯建平
Owner NAT INST FOR FOOD & DRUG CONTROL
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