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Real Time Quantitative PCR Service(qPCR Service)

来源:Seebio.cn作者:西宝生物人气:-发表时间:2012-03-19 13:18:51【

Real Time Quantitative PCR Service(qPCR Service)

Description
Real-time quantitative PCR remains one of the most sensitive and quantitative tools for gene expression used today. Quantitative PCR(qPCR) is a powerful, highly sensitive technique that can be used to quantitate gene expression, determine gene copy number, detect SNPs, and detect DNA from viral and bacterial microorganisms. Seebio support both SybrGreen and Taqman reaction chemistries. With Taqman chemistry, a labeled probe is included in the reaction to enhance the specificity and sensitivity of target sequence detection. With SyberGreen chemistry, amplified product is detected by its interaction with the SyberGreen fluorescent dye, which selectively binds to double-stranded DNA templates. We also offer primer and probe design services. We manage all aspects of the project from assay design to data analysis. Sample preparation (Nucleic acid isolation, whole genome amplification, etc.) is also available. You can get full service from us.

Application
Quantitative PCR can be applied in several key areas including:
1.Gene Expression Analysis
2.SNP Polymorphism Analysis
3.Quantification of miRNA
4.Drug Response Analysis
5.Cell Bank Copy Number Analysis
6.Mutation Analysis
7.Residual DNA Analysis

Reference
1.Chou, Q., Russell, M., Birch, D., Raymond, J. & Bloch, W. (1992) Prevention of pre-PCR misprim­ing and primer dimerization improves low-copy-number amplifications. Nucleic Acids Res.20:1717–1723.
2.Roux, K. H. (1995) Optimization and troubleshooting in PCR. PCR Methods Appl. 4:5185–5194.

3.Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(t)) Method.Methods 2001, 25:402-408.
4.L D Ke, Z Chen .A reliability test of standard-based quantitative PCR: exogenous vs endogenous standards Mol Cell Probes. 2000 Apr;14(2):127-35.
5.Weihong Liu and David A. Saint Validation of a quantitative method for real time PCR kinetics Biochemical and Biophysical Research Communications 294 (2002) 347353
6.Heid, C.A., et al. Real time quantitative PCR. Genome Research 6(10), 986-94, 2002.
7.Pusterla, N., et al. Quantitative real-time PCR for detection of members of the Ehrlichia phagocytophila genogroup in host animals and Ixodes ricinus ticks. J. Clin. Microbiol. 37(5), 1329-31, 1999.
8.Saito T., et al. Quantitative DNA analysis of low-level hepatitis B viremia in two patients with serologically negative chronic hepatitis B. J. Med. Virol. 58(4), 325-31, 1999.
9.Tyagi, S., and F. R. Kramer. Molecular beacons probes that fluoresce upon hybridization. Nature Biotechnology 14(3), 303-8, 1996.
10.Tyagi S., et al. Wavelength-shifting molecular beacons. Nature Biotechnology 18(11), 1191-96, 2000.
11.Park S., et al. Rapid identification of Candida Dubliniensis using a species-specific molecular beacons.J. Clin. Microbiol. 38(8), 2829-36, 2000.

PriceList
Full service (from sample to data acquisition) You can get full service from us

RNA Isolation from Sample

16.00€/sample

cDNA Synthesis

4.00€/reaction

qPCR

12.00/reaction

Taqman Probe

120.00€/probe

Sybr Green I

be free

Time
The turn around time from sample to data is about 30 working days.

Sample Submission Requirements
1.Cells (in vitro cultured cells or white blood cells, purified peripheral blood mononuclear cells (PBMCs): One to 5 million cells. When FACS-sorted and highly purified cells are provided, the number may be as low as 100,000 cells. We have performed TaqMan? analysis for gene transcription using blastocysts at the 32 and 64-cell stage with excellent results.The enough harvested cells which be stored in Trizol? could be shipped frozen (-20°C) in styrofoam boxes.The styrofoam boxes should be recycled.

2. Tissues: Fresh tissue should be provided in a frozen state on dry ice (stored at-80°Cfor up to 1 year). Please provide tissue samples at least 50-100 mg.The grinded sample which be stored in Trizol? could be shipped frozen (-20°C) in styrofoam boxes.The styrofoam boxes should be recycled.

3. Fixed tissues: Two 50 micron sections of paraffin-embedded tissues are required from solid tissues. A third 50 micron section is required from non-solid tissues such as lung tissue. TaqMan? analysis were performed on a variety of paraffin-embedded tissues (i.e. skin, lymph node, GI tract, spleen, liver, lung, brain) which were up to 20 years of age without significant loss of PCR signals for the endogenous control (GAPDH,beta-atin or 18S rRNA) when compared to recently embedded tissue (within 1 year).

 If you want to get more information, please contract us.

Seebio Biotech, Inc.
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