Related Experiment Video
Updated: May 23, 2025
Genotyping of Plant and Animal Samples without Prior DNA Purification
Published on: September 24, 2012
One-tube genotyping using TaqMan-PCR without DNA extraction
Toru Miyagi1,2, Ayano Tahara1, Hironori Kobayashi1
1Japanese Red Cross Kanto-Koshinetsu Block Blood Center, Tokyo, Japan.
Background And Objectives:
Genotyping has become an indispensable technique widely used in transfusion medicine. However, further simplification of the process is essential. Among the simplest methods, TaqMan-PCR is prominent. Nevertheless, challenges related to time, cost and the risk of sample mix-up during the DNA extraction process persist. To address these challenges, we have developed a novel blood processing method.
Materials And Methods:
Whole blood collected from healthy adult blood donors was heated at 95°C for 5 min and then used as a TaqMan-PCR template. The human platelet antigen (HPA)-1, -2, -3, -4, -5 and -15 genotypes of 60 individuals were evaluated.
Results:
The results were fully consistent with those obtained through routine testing using PCR with reverse-sequence-specific oligonucleotide (rSSO) hybridization, confirming the reliability of our approach.
Conclusion:
The developed 'one-tube method' significantly reduces the risk of sample mix-up while offering considerable time and cost savings. Furthermore, this method has broader applicability to other genes, as the polymorphic regions tested in this study exhibited variability in guanine-cytosine content and amplicon lengths.
More Related Videos
05:58Digital Polymerase Chain Reaction Assay for the Genetic Variation in a Sporadic Familial Adenomatous Polyposis Patient Using the Chip-in-a-tube Format
Published on: August 20, 2018
07:00qPCRTag Analysis - A High Throughput, Real Time PCR Assay for Sc2.0 Genotyping
Published on: May 25, 2015
Related Concept Videos
01:24DNA Isolation
02:57Real Time RT-PCR
The real-time quantification of the number of amplified products is...