The Comparison of DNA Quantity between Full and
Half Volume Single Cell Whole Genome Amplification
by Linker-Adapter PCR Technique
Matchuporn Sukprasert MD*,
Sasivimol Rattanasiri PhD**, Kungpu Xu PhD***
Affiliation :
* Reproductive Endocrinology and Infertility Unit, Department of Obstetrics and Gynecology, Faculty of Medicine
Ramathibodi Hospital, Mahidol University, Bangkok, Thailand
** Research Center, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand
*** Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine and Infertility, Weill Cornell Medical
College of Cornell University, New York, USA
Background : Whole genome amplification (WGA) is a very important step in providing sufficient DNA templates for many
high-throughput genetic analyzes. WGA approaches can be subdivided into PCR- or non-PCR-based methods. The PCR
amplification category includes PEP-PCR, DOP-PCR and linker-adapter PCR, but only the linker-adapter PCR is suitable
for application in preimplantation genetic diagnostic screening because it provides the necessary rapid turnaround time.
Objective : Evaluate the ability of linker-adapter WGA commercial kits by using half volume compare with full volume of
the reagent amplified DNA extracted from single cell fibroblast.
Material and Method: Single cell fibroblast was used based on known genetic profiles. The authors reduced the volume of
the reagent and compared the DNA yields and fragmented DNA products with yields and products using the original protocol.
Results : Our result did not show a significant difference between the amount of DNA products between full and half volume
method (4.72 vs. 4.89 μg, p-value = 0.56). We achieve a slightly different of fragmented length of WGA products, full volume
of reagent received slightly longer length than half volume (502.83 vs. 478.30 bp, p-value = 0.19).
Conclusion : In this study, we have shown that the half volume of the reagent of linker-adapter WGA method amplified DNA
extracted from single cell fibroblast was comparable DNA yield and DNA fragmented length with the original method. We
need further study extrapolate to evaluate the outcome.
Keywords : Whole genome amplification, Linker-adapter PCR, DNA yields
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