Detection of homozygous deletion of survival motor neuron 1 (SMN1) gene using allele-specific primer in SMA patients

Hoo, Kit Mei (2007) Detection of homozygous deletion of survival motor neuron 1 (SMN1) gene using allele-specific primer in SMA patients. Universiti Malaysia Sabah. (Unpublished)

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Abstract

This research was carried out to amplify the exon 7 of SMN 1 gene by using allelespecific primer and optimize the peR reagents and parameters for the detection of homozygous deletion of SMNI gene in SMA patients. It had been noticed that 58.4 °c of annealing temperature gave the optimum condition in PCR amplification for both tel-SMN and β-globin primers. Based on the resulted PCR amplification profiles, both SMNI and β-globin genes were performed in the normal healthy sample. Meanwhile, the identification of SMNI gene deletion in SMA patients was verified by the absence of targeted band (SMNI gene) together with the presence of β-globin gene band. This phenomenon could only be detected in the experiment with higher DNA concentration loaded, that was 68 ng. With the β-globin gene served as an internal control, the result was considered valid for any sample that showed β-globin gene band. In order to confirm the actual size of the PCR products, DNA sequencing was performed for both SMNI and β-globin genes. However, the resulted sequencing electropherograms were failed to provide the complete sequence for all samples. Hence, PCR-allele specific method is specifically useful in the analysis of homozygous deletion of SMNI gene in SMA disease but more optimizations are needed for DNA sequencing in order to make this study more reliable and success.

Item Type: Academic Exercise
Uncontrolled Keywords: PCR-allele specific method, DNA, homozygous deletion, temperature
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: SCHOOL > School of Science and Technology
Depositing User: MDM SITI AZIZAH IDRIS
Date Deposited: 02 Apr 2014 02:30
Last Modified: 10 Oct 2017 08:36
URI: http://eprints.ums.edu.my/id/eprint/8018

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