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Real-time PCR Service: Gene Expression Analysis by qPCR

Real-time Quantitative PCR(qPCR)

A real-time quantitative polymerase chain reaction(qPCR) is a technique of molecular biology, that is used to amplify and simultaneously detect or quantify targeted DNA molecules. The procedure follows the general principle of PCR. During PCR reaction progress the accumulation of PCR products results in the enhancement of fluorescence signals as the cycle numbers increase. Thus PCR reaction progress is detected in "real time" by monitoring the fluorescence intensity.

qPCR amplification schematic

qPCR Service Content

qualitative and quantitative gene expression analysis
 
Qualitative Analysis
— detect viruses; detect GMO; identify cultivar; Mutant genotyping
Identification of transgenic mice with human IL6 use SYBR Green qPCR
 
Relative Quantitative Analysis
— reference gene and target gene expression analysis
Reference gene and target gene expression analysis in samples
 
Absolute Quantitative Analysis
— absolute quantitative analysis of DNA(RNA) ; transgene copy determination;
virus and pathogen quantification
Detect the copy number changes of nodavirus after it infect the HighFive cells
 
SNP Analysis
— human disease analysis; Individuation of medical treatment; HRM analysis
Human CA12 gene SNP analysis by HRM

SYBR Green and Taqman Probe qPCR

Quantitative real-time-PCR (qPCR) such as TaqMan and SYBR Green qPCR assays are widely used for gene expression analysis.The advantages and disadvantages of two methods see the table below.
Basic principle of SYBR Green dye method Basic principle of TaqMan probe method

Gene Expression Analysis by qPCR Advantages

Sino biological provide you with SYBR Green dye method and TaqMan probe method for gene expression analysis according to your scientific research demand. Qualitative analysis, relative quantification or absolute quantification can be completed by real-time quantitative PCR.

1.Sino biological has more than 10,000 gene plasmids with high quality and authenticated primer pairs. The detection efficiency is improved greatly.

2. Sino biological also has a number of experienced technical personnels who design qPCR detection schemes and present experimental service.

qPCR Service Process

Taking an example of TaqMan probe method
1. Design and synthesize primers and probes
Customers provide gene information for us, primers and probes are designed by our own algorithm of the company.

2. Perform a real-time PCR experiment
A real-time PCR is performed after RNA in the samples provided by customers is extracted and purified. The examining report is presented.

Examples of qPCR Products

Example 1
The detection results of 45 genes amplified in cDNA of human tissues by SYBR Green qPCR method.
Figure 1. Amplification curves of 45 genes amplified in cDNA of human tissues Figure 2. Melting curves of 45 genes amplified in cDNA of human tissues
 
Example 2
Quality report of one gene primer pairs (including standard curve).
Figure 3. Amplification curves of SDHA standard samples Figure 4. Standard curve and amplification efficiency of SDHA
 
Example 3
qPCR primers of reference genes and standard samples synthesized in our company. Learn more qEASY qPCR primer pairs.
Product Description Catalog#(PDF) Price
Human ACTB qPCR primer pairs HP100001 $79
Human B2M qPCR primer pairs HP100002 $79
Human GAPDH qPCR primer pairs HP100003 $79
Human GUSB qPCR primer pairs HP100004 $79
Human HPRT1 qPCR primer pairs HP100005 $79
Human HSP90AB1 qPCR primer pairs HP100006 $79
Human LDHA qPCR primer pairs HP100007 $79
Human NONO qPCR primer pairs HP100008 $79
Human PGK1 qPCR primer pairs HP100009 $79
Human PPIA qPCR primer pairs HP100010 $79
Human PPIH qPCR primer pairs HP100011 $79
Human RPLP0 qPCR primer pairs HP100012 $79
Human RPLP1 qPCR primer pairs HP100013 $79
Human SDHA qPCR primer pairs HP100014 $79
Human TBP qPCR primer pairs HP100015 $79
Human TFRC qPCR primer pairs HP100016 $79
Figure 5. Quality verification of qPCR primers of part reference genes
The experiment using the cDNA of seven human cancer cells as templates and the corresponding qEASY qPCR primer pairs for SYBR Green quantitative detection.
The expression level of GAPDH is highest and SDHA is most stable.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"