| Topic : Preventive Healthcare: Opportunities & Challenges |
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Moving up the Biotechnology value chain |
Clinical Research - outsourced |
BIOLOGISTICS |
2 more ...
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Source : http://www.expresspharmaonline.com
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last activity : 07 06 2010 20:18:04 +0000
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RT-PCR sets new trends in medical research
The real time Polymerase Chain Reaction (PCR) is faster, less laborious, highly sensitive and specific compared to PCR, and has wide applications
Reverse Transcription Polymerase Chain Reaction (RT-PCR), popularly called 'PCR' is commonly used as a molecular diagnostic tool. In PCR, the appearance of the end product band on an agarose gel takes minimum of 20 to 25 cycles, when the gene amplification is complete as shown in Figure 1. PCR is characterised by high variation and non-reproducibility of results. In real time, the conventional PCR is modified to view the gene expression as it happens. Gene expression is measured at the growth phase, so what you see is the actual amplification. You can see the results as it happens and hence the name real time.
Real time PCR was first documented in 1993. However, the technology came to use only in 2001. It has wide applications and is now used in basic research, molecular medicine, biotechnology and pathological tests. The application of real time PCR is numerous-genomic DNA copy number measurement, mRNA expression, viral DNA, transgene copy number, allelic discrimination assays, microarray data confirmation, analysis of specific splice variants of genes, laser capture micro dissection material, paraffin embedded tissues, flow-sorted cells including stem cells and amplification of RNA from single cell etc. Application in hematology includes measurement of translocation gene product, measurement of DNA copy number in leukemia and expression of gene for differentiation of cell types of an immune response.
Real time PCR is faster, less laborious, highly sensitive and specific compared to the PCR.
There are two types of real time-quantitative and qualitative. In qualitative, the gene expression is with reference to genes such as 18S rRNA or housekeeping gene family like glyceraldehydes-3-phosphate-dehydrogenase (GAPDH) or ß-Actin etc. In quantitative, the exact quantity of the gene expression is measured.
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