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Quantitative Real-Time PCR using the Thermo Scientific Solaris qPCR Assay
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Using synthetic oligonucleotides as standards in probe-based qPCR.

Jillian Conte1,2, Margret J Potoczniak3, Shanan S Tobe3,4

  • 1Keystone College, Department of Biological & Physical Sciences, One College Green, La Plume, PA 18440, USA.

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|April 18, 2018
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Developing DNA standards for quantitative real-time PCR (qPCR) is now faster and safer. Synthetic gene fragments offer a reliable alternative to traditional methods, ensuring comparable assay performance and accuracy for researchers.

Keywords:
DNA standardgBlocks®qPCRsynthetic oligonucleotide

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genomics

Background:

  • Quantitative real-time PCR (qPCR) is a cornerstone technique in molecular biology for DNA quantification.
  • Accurate DNA quantification via qPCR necessitates the use of a standard curve.
  • Traditional standard curve development methods are often laborious, expensive, and carry a risk of contamination.

Purpose of the Study:

  • To evaluate the efficacy of synthetic oligonucleotide gene fragments (gBlocks® Gene Fragments) as an alternative for qPCR standard development.
  • To compare the performance of synthetic gene fragments against purified amplicon standards in qPCR assays.

Main Methods:

  • Utilized gBlocks® Gene Fragments as a standard for DNA quantification in qPCR.
  • Compared the sensitivity, reliability, and overall assay performance using synthetic standards versus purified amplicon standards.

Main Results:

  • gBlocks® Gene Fragments demonstrated comparable sensitivity to purified amplicon standards.
  • The use of synthetic gene fragments resulted in reliable qPCR assay performance.
  • Synthetic standards provided an accurate solution for DNA quantification, overcoming drawbacks of traditional methods.

Conclusions:

  • Synthetic oligonucleotide gene fragments are a viable and effective alternative for qPCR standard development.
  • Researchers can achieve accurate and reliable DNA quantification using synthetic standards, saving time and reducing contamination risks.