Applications of homemade kit in mutation detection of genes |
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Authors: | Chunxia?Zhao Email author" target="_blank">Guowang?XuEmail author Xianzhe?Shi Jianmei?Ma Yan?Zhang Shen?Lü Qing?Yang |
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Institution: | 1. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011, China 2. Cell Laboratory, Second Affiliated Hospital, Dalian Medical University, Dalian 116023, China 3. Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011, China; Cell Laboratory, Second Affiliated Hospital, Dalian Medical University, Dalian 116023, China |
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Abstract: | Several methods of mutation detection, such as single-strand conformation polymorphism (SSCP), tandem SSCP/heteroduplex analysis
and SNaPshot analysis were developed using homemade kit on AB1 310 genetic analyzer, and were successfully applied to mutation
detection of 31 colorectal tumor samples. The sieving capability of homemade kit and commercial kit were compared, results
demonstrate that homemade kit has higher resolution and shorter analysis time. In clinical tumor samples, 26% K-ras (exon
1) and 24% p53 (exons 7–8) were found to have mutations, and all mutations were single point variations. A majority of mutations
occurred in one gene, only 1 tumor contained alterations in the two genes, which indicates that development of colorectal
cancer lies on alternate pathways, and may correlate with different gene mutations |
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Keywords: | single nucleotide polymorphism (SNP) single-strand conformation polymorphism (SSCP) heteroduplex analysis (HA) SNaPshot linear polyacrylamide (LPA) polydimethylacrylamide (PDMA) |
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