• Journal of Genetic Medicine
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• v.9 no.1
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• pp.11-16
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• 2012

Purpose: To determine a method to improve the efficacy and accuracy of preimplantation genetic diagnosis (PGD) - polymerase chain reaction (PCR), we compared hot start PCR and conventional multiplex nested PCR. Materials and Methods: This study was performed with single lymphocyte isolated from whole blood samples that were obtained from two couples with osteogenesis imperfecta (OI). We proceeded with conventional multiplex nested PCR and hot start PCR in which essential reaction components were physically removed, and we compared the amplification rate, allele dropout rate and nonspecific products. Afterward, we used selective method for PGD. Results: In the two couples, the respective amplification rate were 93.5% and 80.0% using conventional multiplex nested PCR and 95.5% and 92.0% using hot start PCR. The respective mean allele dropout rates for the two couples were 42.0% and 14.0% with conventional multiplex nested PCR and 36.0% and 6.0% with hot start PCR. Conclusion: The results demonstrate that the hot start PCR procedure provides higher amplification rates and lower allele dropout rate than the conventional method and that it decreased the nonspecific band in multiplex nested PCR. The hot start method is more efficient for analyzing a single blastomere in clinical PGD.

• Korean Journal of Veterinary Service
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• v.33 no.4
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• pp.361-366
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• 2010

This study was carried out to investigate the characters of short-tailed dogs (DongGyeong dogs) with anatomical insights, molecular genetics in Gyeongju. The present study was conducted to further characterize of short-tailed dog population in Gyeongju. The short-tailed dog was analyzed in the distribution of 55 individual. The anatomical insights were by x-ray. For discovery of specific genes expressions were measured by Hot-start PCR analysis. Anatomy survey, the number of vertebral typical consists of more than 20. 88.9% of short-tailed dog populations consists of 3-8 vertebrates. The 54 individuals of the 47 observe the vestigial tailed of the sacrum. No detected sacrococcygeal vertebrae degradation individuals were malformation defects. The 3 genes were DEGs (differentially expressed genes) in Dong-Gyeong dogs. We succeeded in finding 3 novel DongGyeong dogs specific genes by using Hot-start PCR analysis, this study suggests that these novel genes may play role (s) in DongGyeong dogs.

• Journal of Oral Medicine and Pain
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• v.19 no.2
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• pp.205-219
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• 1994

Cigarette butts from 5 smokers were gathered and then, placed in room temperature for 1, 3, 5, 7, 15 days. The possible use of the cigarette butts for individual identification was evaluated in sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene from the extracted DNA. 1. DNA extraction was possible in cigarette butts weree left in room temperature for 15days, so it can be applicatable to individual identification by polymerase chain reaction(PCR). 2. Amplification of X-Y homologous amelogenin gene by PCR made it possible to identify the sex in saliva stains (cigarette butts). 3. Amplification of D1S80 locus can be acquired from adding the boving serum albumin and hot start PCR procedures from forensic samples such as saliva stains (cigarette butts), so the AMP-FLPs examining is possible. 4. Genotype could be determined simply and rapidly using Amplitype$TM$ HLA-DQ$\alpha$ forensic kit in examining the HLA-DQA1 gene. From the investigation, DNA extraction, sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene was successfully done even though the cigarette butts were left for 15 days at room temperature. Therefore cigarette butts are highly reliable and applicatable as molecular biologic samples for individual identification.