Composite amplification system and detection kit of mouse short tandem repeat

A compound amplification system and short tandem repeat technology, applied in the biological field, can solve the problem that STR typing has not been widely used

Active Publication Date: 2012-07-11
SUZHOU MICROREAD GENETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for the identification of mouse cell lines, STR typing has not been widely used because there is no established multiple STR amplification system and related products

Method used

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  • Composite amplification system and detection kit of mouse short tandem repeat
  • Composite amplification system and detection kit of mouse short tandem repeat
  • Composite amplification system and detection kit of mouse short tandem repeat

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Experimental program
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Effect test

Embodiment 1

[0106] The following is our application of the present invention to 15 cases of mouse cells CTLL-2, 3T6 Swiss, NS-1, P3X63Ag8.65, J774A.1, LM(TK-), WEHI-13VAR, P388, SP2 / 0, SC-1 , BALB / C 3T3Clone A31, M.dunni, NIH / 3T3, A9 and 3T3 Swiss DNA samples for the specific implementation of testing. The samples are all common cells and are commercially available. Our company also keeps them and can be publicly distributed.

[0107] 1. DNA extraction

[0108] Tissue DNA extraction kit (Tiangen Biochemical) was used to analyze the cultured CTLL-2, 3T6 Swiss, NS-1, P3X63Ag8.65, J774A.1, LM (TK-), WEHI-13VAR, P388, SP2 / 0, SC -1. BALB / C 3T3Clone A31, M. dunni, NIH / 3T3, A9 and 3T3 Swiss cells were subjected to DNA extraction. The operation steps were in accordance with the instructions. After DNA extraction, the DNA was quantified with an ultraviolet spectrophotometer and diluted to 2ng / μl.

[0109] 2. PCR

[0110] 2.1 Reaction system:

[0111] A total of 8 pairs of primers at Jarid1 site and 7 STR...

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Abstract

The invention relates to a composite amplification system and a detection kit of a mouse short tandem repeat and belongs to the technical field of biotechnology. The invention relates to mouse short tandem repeats (D2Mit395.1, D4Mit170.1, D5Mit201.1, D7Mit40, D11Mit4, D15Mit16 and D17Mit51.1) and a sex determination site (Jarid1) for simultaneously amplifying mice in a PCR (polymerase chain reaction) system, carrying out composite amplification and capillary electrophoresis detection on mouse DNA (deoxyribonucleic acid). The system can be used for genotyping of a mouse cell, each site adopts fluorescence-labeled primers, so that an obtain product is provided with a fluorescence label, and the detection can be carried out on an apparatus such as a genetic analyzer, so that the obtain segment length is more exact, and higher in repeatability. The system is simple to operate and convenient for large-scale popularization, the whole process has automation equipment, and the whole detectiontime only needs about 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 quickly and accurately perform STR typing of mouse cells and then identify the types of mouse cell lines. Background technique [0002] In 2009, an editorial on Nature pointed out: Among the thousands of biological laboratories that use cell lines, many laboratories do not know that commonly used cell lines between 1 / 5 and 1 / 3 may no longer be used. It was originally thought. In the past 25 years, a large number of studies, together with the experience of cell banks in the United States, the United Kingdom, Germany, and Japan, found that 18-36% of cultures contained misidentified species or other cell types. Even the cell bank of the National Cancer Institute, which is under strict management, has been contaminated with cell lines. Now the only way for laboratory res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 陈初光周仲春吕悦心
Owner SUZHOU MICROREAD GENETICS
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