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Related Experiment Videos

Multiple displacement amplification on single cell and possible PGD applications.

Ali Hellani1, Serdar Coskun, Moncef Benkhalifa

  • 1King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia. hellani@kfshrc.edu.sa

Molecular Human Reproduction
|October 7, 2004
PubMed
Summary
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Multiple displacement amplification (MDA) enables whole genome amplification from single cells, improving upon PCR and FISH for preimplantation genetic diagnosis (PGD). This technique shows promise for single-cell molecular karyotyping and genetic disorder diagnosis.

Area of Science:

  • Genetics
  • Molecular Biology
  • Reproductive Medicine

Background:

  • Preimplantation genetic diagnosis (PGD) traditionally relies on PCR and FISH, which have limitations in DNA quality and chromosome diagnosis.
  • Whole genome amplification from single cells is crucial for comprehensive genetic analysis in PGD.

Purpose of the Study:

  • To evaluate Multiple Displacement Amplification (MDA) for single-cell whole genome amplification.
  • To assess the accuracy and utility of MDA for PGD applications.

Main Methods:

  • Whole genome amplification of DNA from single cells using MDA.
  • Analysis of 16 short tandem repeats (STRs) to assess amplification patterns.
  • Evaluation of allelic dropout (ADO) using beta globin gene sequencing and STR analysis.

Related Experiment Videos

  • Detection of trisomy 21 using comparative genome hybridization-array.

    • Main Results:

    • MDA successfully amplified 16 STRs with patterns similar to genomic DNA.
    • Allelic dropout rates were 10.25% for beta globin gene sequencing and 5% for STR analysis.
    • MDA enabled accurate detection of trisomy 21 in a single cell.

    Conclusions:

    • MDA is a viable method for single-cell whole genome amplification.
    • MDA shows potential for single-cell molecular karyotyping and diagnosing single gene disorders in PGD.