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

• 발행기관: 대한의학유전학회
• 저자명: Sung Ah Kim Moon Joo Kang Hee Sun Kim Sun Kyung Oh Seung-yup Ku Young Min Choi Jong Kwan Jun Shin Yong Moon
• 간행물 정보: 『대한의학유전학회지』제9권 제1호, 11~16쪽, 전체 6쪽
• 주제분류: 의약학 > 기타의약학
• 파일형태: PDF
• 발행일자: 2012.09.01

영문 초록

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.

목차

Introduction

Materials and Methods

Results

Discussion

Acknowledgment

Reference

키워드

Hot start PCR Nested PCR Preimplantation genetic diagnosis