Kahar, Nazirah Abd
(2009)
Investigation of CTCF gene in Glioma and its
correlation with YB-1 gene using SYBR-Green I based
real-time PCR.
Project Report.
Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia.
(Submitted)
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 identifying 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 ΔΔCt value determination. Comparative
method of ΔΔCt 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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