Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Primer, kit and detection method used for detecting insect cell DNA residues

A technology of insect cells and kits, applied in the biological field, can solve the problem of no rapid quantitative detection kits for DNA residues in insect cells, and achieve reliable quality detection data, good sensitivity and specificity

Inactive Publication Date: 2020-12-18
天津科佰迪生物医药科技有限公司
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although real-time fluorescent PCR technology (dye method) has been used to detect residual DNA in CHO cells, there is currently no rapid quantitative detection kit for residual DNA in insect cells in the detection of residual DNA in vaccines and gene therapy products.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Primer, kit and detection method used for detecting insect cell DNA residues
  • Primer, kit and detection method used for detecting insect cell DNA residues
  • Primer, kit and detection method used for detecting insect cell DNA residues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1.1 Materials and reagents

[0036] 2×Taq PCR master mix was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.;

[0037] 2 × Taqman Master Mix premix, purchased from Tianjin Kebaidi Biomedical Technology Co., Ltd.;

[0038] ABI7500 fluorescent quantitative PCR instrument, purchased from ABI (Applied Biosystems, USA);

[0039] The DNA diluent is prepared by the laboratory using molecular biology grade reagents;

[0040] Genomic DNA extraction kit was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.;

[0041] The primers and probes used in the experiment were synthesized and purified by Invitrogen American Yingjie Life Technology Co., Ltd.

[0042] The primer and probe sequences used are as follows:

[0043] Forward primer sequence: 5'- CGCTGTAAGAGGACCGTAG -3' (Seq ID No:1)

[0044] Reverse primer sequence: 5'- GACCTCACGCAATCTTCTGA -3' (Seq ID No:2)

[0045] Taqman probe: 5'- ACCGCTGCCTCCTCCTGCAGG -3' (Seq ID No:3)

[0046] The fluo...

Embodiment 2

[0068] Embodiment 2: the preparation of kit

[0069] The preparation of the kit includes the following components:

[0070] Insect cell genomic DNA (30ng / ul) (40ul / tube) 1 tube, DNA diluent (7ml / bottle) 1 bottle, 2×TaqmanMaster Mix (1500ul / tube) 1 tube, 10×primer-probe mixture (300ul / tube ) 1 tube, ultrapure water (1.5ml / tube) 1 tube.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a primer, a kit and a detection method for detecting insect cell DNA residues. Through a method of using a real-time fluorescent quantitative PCR technology for qualitatively and quantitatively detecting the insect cell DNA residues and a PCR kit prepared according to the method, a quantitative limit of as low as 3fg / ul for detecting the insect cell DNA residues is realized; and in other words, the method has good sensitivity and specificity. The real-time fluorescent quantitative PCR method can be used for detecting recombinant protein or recombinant virus vaccines orgene therapy vectors produced by insect cells which are taken as expression host cells, such as recombinant adeno-associated viruses and the like, and can provide reliable quality detection data for research, development and safe production of the gene therapy vectors.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a primer, a kit and a detection method for detecting insect cell DNA residues. Background technique [0002] Real-time fluorescent PCR technology is a nucleic acid detection technology that has developed rapidly in recent years. It reflects the amplification level of each cycle of PCR in real time by detecting the dynamic changes of fluorescent signals. The CCD can periodically emit excitation light of a specific wavelength to generate excitation light, and then the CCD collects the signal of the excitation light, and analyzes and summarizes it to the workstation through software to obtain the amplification curve. TaqMan PCR technology is a kind of real-time fluorescent PCR. Compared with the traditional PCR, it adds a probe whose two ends are respectively labeled with a fluorescent group and a quencher group in the reaction system. When the probe structure is complete, the fluores...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/6888C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6888C12Q2531/113C12Q2561/101
Inventor 陈涛王博玮李雅雯
Owner 天津科佰迪生物医药科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com