Unlock instant, AI-driven research and patent intelligence for your innovation.

A combination of qPCR internal reference genes in lung tissue of a mouse model of kidney yang deficiency and exogenous infection and its screening method

A technology of internal reference gene and mouse model, applied in the field of molecular biology

Active Publication Date: 2022-06-24
SHANDONG UNIV OF TRADITIONAL CHINESE MEDICINE
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, the literature is more common in Magnolia [8] Alisma [9] , Chuanduan [10] There are more and more reports on RT-qPCR internal reference gene screening of medicinal plants, but there is no report on RT-qPCR internal reference gene screening of lung tissue of kidney-yang deficiency exogenous mouse model

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
  • A combination of qPCR internal reference genes in lung tissue of a mouse model of kidney yang deficiency and exogenous infection and its screening method
  • A combination of qPCR internal reference genes in lung tissue of a mouse model of kidney yang deficiency and exogenous infection and its screening method
  • A combination of qPCR internal reference genes in lung tissue of a mouse model of kidney yang deficiency and exogenous infection and its screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] 1 Materials and methods

[0033] 1.1 Animals

[0034] SPF grade Balb / c male mice, weighing 18-20 g, were purchased from Jinan Pengyue Laboratory Animal Breeding Co., Ltd., license number: SCXK (Lu) 20140007.

[0035] 1.2 Reagents

[0036] Estradiol benzoate (batch number: 140627, purchased from Ningbo No. 2 Hormone Factory, 2 mg / mL), H1N1 influenza virus (Beijing local strain cited from China CDC Institute of Virology), 0.9% sodium chloride injection (Chenxin Pharmaceutical Co., Ltd., batch number: 1601042161), RNAstore sample preservation solution-DP408, animal tissue total RNA extraction kit-DP431 (RNAprepPure Tissue Kit), cDNA reverse transcription kit-KR106, SYBR Green fluorescence quantitative PCR kit- FP205 (Tiangen Company).

[0037] 1.3 Instruments

[0038] Quawell ultra-trace UV spectrophotometer (USA), fluorescence quantitative PCR instrument (CFX Connect Real-TimeSystem, Bole), rectal temperature tester: BAT-12 microprobe thermometer (Physitemp Instrument...

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
PCR efficiencyaaaaaaaaaa
Login to View More

Abstract

The present invention provides a method for screening the combination of qPCR internal reference genes in the lung tissue of the kidney-yang deficiency exogenous mouse model, designing 15 internal reference genes Actb, β2m, Gapdh, Gusb, Tuba, Grcc10, Eif4h, Rnf187, Nedd8, Ywhea, 18S rRNA , RPL13, Ubc, RPL32, the specific primers of Ppia, the sequence of the upstream and downstream primers is shown in SEQ ID NO.1‑30, and then using the cDNA of the kidney-yang deficiency exogenous mouse model as a template, using the specific primer pair respectively The RT-qPCR analysis of 15 internal reference genes, combined with the t-test between groups and the absolute value comparison of the average values ​​between groups, can conveniently and quickly establish the qPCR internal reference gene combinations of the kidney-yang deficiency exogenous mouse model lung tissue as Grcc10 and Ppia.

Description

technical field [0001] The invention relates to molecular biology technology, in particular to an internal reference gene combination based on qPCR technology and a screening method thereof. Background technique [0002] Quantitative real-time PCR (qPCR) is a quantitative method for real-time monitoring and analysis of PCR amplification products. It has many advantages such as high sensitivity, good repeatability, fast detection speed, and high degree of automation. It has been widely used by scholars at home and abroad. This technique generally requires the correction and normalization of the expression of the target gene with a stable internal reference gene. The ideal reference gene should be constantly expressed in various types of tissues and cells under various experimental conditions, and its expression levels should be similar without significant differences. However, the transcription levels of reference genes may change in different types of tissues, developmenta...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/166C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 杨勇容蓉付业佩杨佳范珊珊赵绍哲
Owner SHANDONG UNIV OF TRADITIONAL CHINESE MEDICINE