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

T2DM (Type 2 Diabetes Mellitus) detection primer group, PCR (Polymerase Chain Reaction) chip and detection method for human and monkeys

A type 2 diabetes detection chip technology, applied in the field of fluorescent quantitative PCR detection chip, can solve the problems of failure to obtain primer sets and other problems

Inactive Publication Date: 2013-03-13
广东蓝岛生物技术有限公司
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no fluorescent quantitative PCR chip that can simultaneously analyze the expression status of T2DM-related genes in humans and cynomolgus monkeys. The main difficulty lies in the lack of primer sets that can be amplified under the same conditions and used to analyze the expression of T2DM-related genes in human and cynomolgus monkeys.

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
  • T2DM (Type 2 Diabetes Mellitus) detection primer group, PCR (Polymerase Chain Reaction) chip and detection method for human and monkeys
  • T2DM (Type 2 Diabetes Mellitus) detection primer group, PCR (Polymerase Chain Reaction) chip and detection method for human and monkeys
  • T2DM (Type 2 Diabetes Mellitus) detection primer group, PCR (Polymerase Chain Reaction) chip and detection method for human and monkeys

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1. Extraction of white blood cells.

[0058] ① Collect venous blood in the anticoagulant tube and mark each other; the samples 1-5, 26-32 health monkey samples; the sample 6-10 is the crab monkey sample of the initial stage of the model; the sample 11-15 is the model of the crab monkeys in the model;Sample 16-20 clinical early clinical crab and monkey sample; sample 21-25, 33-39 is a clinical crab monkey sample; sample 40-46 is a healthy sample; sample 47-53 is a diabetic sample.

[0059] ② Add 1ml lymphocyte extract to the 25ml centrifugal tube.

[0060] ③ Slowly add an equal volume of PBS buffer (1 ×) to the anticoagulant tube equipped with 10ml blood, mix well.

[0061] ④ Slowly add the mixture of ③ ② to ② with a pipetter, and try not to sink the mixed liquid into the bottom of the ②, causing it to suspend it on the lymphocyte separation liquid.

[0062] ⑤ Corporation on frozen centrifuge, 4 ℃, 1700g, 15min.

[0063] ⑥ Take out the centrifugal tube and carefully ...

Embodiment 2

[0066] Example 2. Extract of RNA.

[0067] ① Take out the leukocytes obtained from the implementation example 1 from the liquid nitrogen. The room temperature is static. The finger wall of the bomb pipe can be scattered. The 1ml Trizol is added to each frozen tube to fully mix.

[0068] ② Layers: After the sample is added to Trizol, the room temperature is placed for 5 minutes to make the sample split.4 ° C, 15000g centrifugal 10min take the Qing.Add 200 μL of chloroform in Shangqing. After the severe oscillation is mixed, the room temperature is placed for 3-5min.

[0069] ③ RNA precipitation: 4 ℃, 15000g centrifugal 10-15min.The sample will be divided into three layers: yellow organic phase, the middle layer and colorless water phase, and the RNA is mainly in the water phase, and the water phase (usually 550 μL can be absorbed) to the new tube.Be careful to absorb the water phase. Do not absorb the intermediate interface, otherwise it will cause DNA pollution in RNA samples.Add ...

Embodiment 3

[0072] Example 3.CDNA reverse transcription.

[0073] Use Transgene's CDNA transliteration kit-Easyscript First-Strand CDNA SYNTHESIS SUPERMIX (TRANSGEN BIOTECH, AE301) to transcrIn the system, the total RNA volume is about 0.25-0.5 μg.

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

PropertyMeasurementUnit
Upstream primeraaaaaaaaaa
Login to View More

Abstract

The invention relates to a PCR (Polymerase Chain Reaction) primer group, a fluorescence quantitative PCR detection chip and a detection method for analyzing the associated gene expression state of T2DM (Type 2 Diabetes Mellitus) for human and Cynomolgus monkeys simultaneously. The T2DM detection primer group for human and monkeys comprises amplification primer pairs respectively for amplifying T2DM disease associated protein genes as follows: ACE, ACLY, G6PD, GSK3B, HMOX1, IDE, PRKCB1, PYGL, AQP, CCR2B, CEACAM1, CTLA4, GCGR, ICAM1, NSF, RAB4A, SELL, SNAP23B, STX4, STXBP2, TNFRSF1A, VAMP3, VAPA, IFNG, INS, TGFB1, TNFA, IGFBP5, INPPL1, PIK3C2B, PIK3R1, IKBA, NEUROD1, NFKB1, PPARGC1A and AGT, and the sequences of the gene amplification primer pairs are nucleotide sequences respectively shown as SEQ ID NO: 1-72. The invention also provides the fluorescence quantitative PCR detection chip and the detection method comprising the PCR primer group. The invention has the beneficial effects of performing accurate quantification and detection on T2DM disease associated genes for human and monkeys simultaneously, thus having far reaching importance for promoting fundamental research, preventive detection and clinical treatment of diabetes.

Description

Technical field [0001] The invention involves the PCR primer group, fluorescent quantitative PCR detection chip and detection method for analyzing the gene expression state of human and crab monkeys. Background technique [0002] Type 2 diabetes (referred to as T2DM) is a type of diabetes, also known as non -insulin dependent diabetes.The incidence of diabetes foot, eye lesions, kidney lesions, neuropathy, etc.) and high mortality rates of disability and death; used to be considered a middle -aged and elderly disease. In recent yearsThe disease is young. [0003] In recent years, with the improvement of people's living standards, the incidence of diabetes is rising year by year. Diabetes has become the third non -infectious disease that seriously endangers human health after cardiovascular and cerebrovascular diseases and tumors. The prevention and treatment of diabetes has becomeAn important topic of scientific workers.T2DM is caused by the common role of genetic and environment...

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): C12Q1/68C12N15/11
Inventor 彭白露刘晓明季芳郝香芬夏机良曾小明
Owner 广东蓝岛生物技术有限公司
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