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Different denaturation rates between methylated and non-methylated genomic DNA can result in allele-specific PCR amplification

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Open Journal of Genetics 2011年第2期1卷 13-14页

作者： David J. Bunyan Hilary M. S. Bullman Margaret Lever Sasi D. Saminathan wee teik keng Roziana Araffin David O. Robinson 不详

We analysed a DNA sample from a father and child who were both heterozygous for a 7 base pair insertion in the MEST gene differentially-methylated promoter region, previously shown by PCR analysis of bisulphite-treated DNA to be on the methylated allele in the unaffected father and the unmethylated allele in the affected child. PCR from genomic DNA was then carried out using a commercial PCR kit with its recommended initial DNA denaturation step of 2 minutes. Subsequent sequence analysis showed that only the non-methylated allele had been amplified, the father appearing to be homozygous normal and the child appearing to have a homozygous 7 b.p. insertion. The PCR protocol was then modified in order to use a longer DNA denaturation stage prior to the addition of the polymerase enzyme. Upon doing so, both the methylated and non-methylated alleles were then identifiable by sequencing with the mutation appearing in its expected heterozygous form. These results highlight the fact that the methylation status of DNA can affect the denaturation rate prior to PCR and result in allele drop-out, showing that the standard protocols of commercial kits should be used with caution when working with methylated regions of DNA.

关键词： Differential Methylation Allele-Specific PCR Commercial Kit

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Polygon Nesting Using Genetic Algorithm

Polygon Nesting Using Genetic Algorithm

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第五届ABB杯全国自动化系统工程师论文大赛

作者： Chen Chen Chua Kim Huat wee keng Hwee Sembcorp Marine Technology Pte. National Vniversity of Singapore.

The nesting problem has been for many years a subject of interest to shipbuilders as continuously rising price of raw *** optimized steel plate cutting plan saves shipyard both time and material,improves production efficiency,and in return enhances the company’s competition *** this paper,the author will propose a polygon nesting method using bottom left strategy with genetic *** algorithm allows parts to be nested within several steel plates which have fixed sizes by searching a suitable accommodate node among the intersection points of the no-fit polygons of previously placed parts relative to the new part following a bottom left *** verify this algorithm using a rectangular nesting problem from published *** then give more computational experiments on polygon nesting problems.

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Polygon Nesting Using Genetic Algorithm

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自动化博览 2011年第S2期28卷 43-48页

作者： Chen Chen~1 Chua Kim Huat~2 wee keng Hwee~3 (1.Sembcorp Marine Technology Pte.) (2.National Vniversity of Singapore.) (3.Sembcorp Marine Technology Pte.)

关键词：自动化系统工程师增刊论文集

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