Application of Hot Start PCR Method in PCR-based Preimplantation Genetic Diagnosis

Kim, Sung-Ah; Kang, Moon-Joo; Kim, Hee-Sun; Oh, Sun-Kyung; Ku, Seung-Yup; Choi, Young-Min; Jun, Jong-Kwan; Moon, Shin-Yong

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자료유형: 학술저널

저자정보: Kim, Sung-Ah (Department of Obstetrics and Gynecology, Seoul National University Hospital) Kang, Moon-Joo (Department of Obstetrics and Gynecology, Seoul National University Hospital) Kim, Hee-Sun (Department of Obstetrics and Gynecology, Seoul National University Hospital) Oh, Sun-Kyung (Department of Obstetrics and Gynecology, Seoul National University Hospital) Ku, Seung-Yup (Department of Obstetrics and Gynecology, Seoul National University Hospital) Choi, Young-Min (Department of Obstetrics and Gynecology, Seoul National University Hospital) Jun, Jong-Kwan (Department of Obstetrics and Gynecology, Seoul National University Hospital) Moon, Shin-Yong (Department of Obstetrics and Gynecology, Seoul National University Hospital)

저널정보: 대한의학유전학회 Journal of genetic medicine Journal of genetic medicine 제9권 제1호

발행연도: 2012.1

수록면: 11 - 16 (6page)

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2012

Application of Hot Start PCR Method in PCR-based Preimplantation Genetic Diagnosis

Kim , Sung-Ah , Kang 외 13명 Journal of genetic medicine 2012.01 학술저널

2007

Investigation of Duplex-nested PCR and Triplex-nested PCR for Preimplantation Genetic Diagnosis of Duchenne Muscular Dystrophy (DMD)

Kim , Sung-Ah , Yoon 외 17명 대한생식의학회 학술대회 2007.01 학술대회자료

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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.

#Hot start PCR #Nested PCR #Preimplantation genetic diagnosis

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참고문헌 신청

Lebedev A / 2009 / Heat-activatable primers for hot-start PCR: oligonucleotide synthesis and basic PCR setup / Curr Protoc Nucleic Acid Chem 4(35):1~17