Application of Hot Start PCR Method in PCR-based Preimplantation Genetic Diagnosis
Journal of Genetic Medicine
;
: 11-16, 2012.
Article
in English
| WPRIM
| ID: wpr-66747
ABSTRACT
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 ANDMETHODS:
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Osteogenesis Imperfecta
/
Patient Dropouts
/
Prostaglandins D
/
Blastomeres
/
Lymphocytes
/
Family Characteristics
/
Polymerase Chain Reaction
/
Preimplantation Diagnosis
/
Alleles
Type of study:
Diagnostic study
Limits:
Humans
Language:
English
Journal:
Journal of Genetic Medicine
Year:
2012
Type:
Article
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