Publication: Investigation of ctcf gene in glioma and its correlation with yb-1 gene using sybr-green i based real-time pcr.
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Date
2009
Authors
Kahar, Nazirah Abd
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Abstract
A glioma brain tumor is a primary brain tumor that originates from the supportive cells of the brain, known as glial cells consists of astrocytes, oligodendrocytes and ependymal. Unlike neurons, these cells have the ability to divide and multiply. Though very rare, mostly they are malignant when occurs. These malignancies pose the greatest clinical problems due to difficulty in diagnosis. Even though scans are done, it only produces pictures that suggest a particular type of tumor. The definitive diagnosis is via sample biopsy of tumor examined under a microscope by neuropathologist. Reliable genetic markers are urgently needed to identify glioma patients to avoid invasive approach. The first step in identifiying genetic marker is to analyze the gene expression in the patient sample in comparison to normal sample either its being up-regulated or down-regulated. Therefore, Total RNA is extracted and converted to eDNA and subsequently act as template in PCR providing Ct value by using MCt value determination. Comparative method of MCt provides the data of fold change which is useful to screen genes either being up-regulated or down-regulated in the patients in comparison to non-cancerous normal sample. However, the analysis expression of selected genes is unable to be accessed as the Ct value is unacceptable based on the follow-up assays for amplicon identification in PCR. Instead, the guidelines to optimize the result were obtained based on the experiences and flaws encountered during the experiment. Thus, guidelines reported here are very crucial to be followed by others who interested in gene expression experiment in the future so that good result can be produced.
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Keywords
glioma brain tumor