J Med Assoc Thai 2013; 96 (11):1491

Views: 1,140 | Downloads: 28 | Responses: 0

PDF XML Respond to this article Print Alert & updates Request permissions Email to a friend

The Comparison of DNA Quantity between Full and Half Volume Single Cell Whole Genome Amplification by Linker-Adapter PCR Technique
Sukprasert M Mail, Rattanasiri S , Xu K

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

Download: PDF