Investigation of CTCF gene in Glioma and its correlation with YB-1 gene using SYBR-Green I based real-time PCR

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...

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Main Author: Kahar, Nazirah Abd
Format: Monograph
Language:English
Published: Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia 2009
Subjects:
Online Access:http://eprints.usm.my/59411/
http://eprints.usm.my/59411/1/NAZIRAH%20BINTI%20ABD.%20KAHAR%20-%20e.pdf
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author Kahar, Nazirah Abd
author_facet Kahar, Nazirah Abd
author_sort Kahar, Nazirah Abd
building USM Institutional Repository
collection Online Access
description 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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format Monograph
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institution Universiti Sains Malaysia
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language English
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publishDate 2009
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spelling usm-594112023-11-14T04:34:59Z http://eprints.usm.my/59411/ Investigation of CTCF gene in Glioma and its correlation with YB-1 gene using SYBR-Green I based real-time PCR Kahar, Nazirah Abd RC31-1245 Internal medicine 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. Pusat Pengajian Sains Kesihatan, Universiti Sains Malaysia 2009 Monograph NonPeerReviewed application/pdf en http://eprints.usm.my/59411/1/NAZIRAH%20BINTI%20ABD.%20KAHAR%20-%20e.pdf 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)
spellingShingle RC31-1245 Internal medicine
Kahar, Nazirah Abd
Investigation of CTCF gene in Glioma and its correlation with YB-1 gene using SYBR-Green I based real-time PCR
title Investigation of CTCF gene in Glioma and its correlation with YB-1 gene using SYBR-Green I based real-time PCR
title_full Investigation of CTCF gene in Glioma and its correlation with YB-1 gene using SYBR-Green I based real-time PCR
title_fullStr Investigation of CTCF gene in Glioma and its correlation with YB-1 gene using SYBR-Green I based real-time PCR
title_full_unstemmed Investigation of CTCF gene in Glioma and its correlation with YB-1 gene using SYBR-Green I based real-time PCR
title_short Investigation of CTCF gene in Glioma and its correlation with YB-1 gene using SYBR-Green I based real-time PCR
title_sort investigation of ctcf gene in glioma and its correlation with yb-1 gene using sybr-green i based real-time pcr
topic RC31-1245 Internal medicine
url http://eprints.usm.my/59411/
http://eprints.usm.my/59411/1/NAZIRAH%20BINTI%20ABD.%20KAHAR%20-%20e.pdf