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Related Concept Videos

Targeted Cancer Therapies02:57

Targeted Cancer Therapies

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The targeted cancer therapies, also known as “molecular targeted therapies,” take advantage of the molecular and genetic differences between the cancer cells and the normal cells. It needs a thorough understanding of the cancer cells to develop drugs that can target specific molecular aspects that drive the growth, progression, and spread of cancer cells without affecting the growth and survival of other normal cells in the body.
There are several types of targeted therapies against...
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Related Experiment Video

Updated: Apr 5, 2026

Employing Digital Droplet PCR to Detect BRAF V600E Mutations in Formalin-fixed Paraffin-embedded Reference Standard Cell Lines
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Detection of BRAF mutations from solid tumors using Tumorplex™ technology.

Jacob Yo1, Katie S L Hay1, Dilanthi Vinayagamoorthy1

  • 1MultiGEN Diagnostics LLC, 854 Paragon Way, Rock Hill, SC 29730, United States.

Methodsx
|August 11, 2015
PubMed
Summary
This summary is machine-generated.

Tumorplex™ is a novel allele-specific sequencing method for detecting single-base mutations in tumor biopsies. This highly sensitive assay identifies mutant alleles without suppression by wild-type signals, even in small or heterogeneous samples.

Keywords:
BRAF V600EFormalin fixed paraffin embeddedLowest level of detectionMultiplex Sanger sequencing platformTumorplex™TumorsWeighted sequencing primers

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Area of Science:

  • Molecular Biology
  • Genetics
  • Oncology

Background:

  • Allele-specific multiplex sequencing (Tumorplex™) represents a significant advancement in molecular diagnostics.
  • Detecting single-base mutations in tumor biopsies is crucial for accurate clinical testing and personalized medicine.

Purpose of the Study:

  • To introduce and evaluate the Tumorplex™ platform for sensitive detection of single-base mutations.
  • To demonstrate the capability of Tumorplex™ in identifying mutant alleles without wild-type signal suppression.

Main Methods:

  • Tumorplex™ utilizes a modified Sanger sequencing approach with differentially weighted primers.
  • Simultaneous generation of mutant and wild-type sequences in a single electropherogram.
  • Assay validation using BRAF V600E mutation detection in cloned DNA and tumor cell lines.

Main Results:

  • The Tumorplex™ platform demonstrated high sensitivity, with a lower limit of detection of 20 genome equivalents (GE) for BRAF V600E.
  • The assay successfully detected mutant BRAF signals at a mutant to wild-type allele ratio of 20:1 × 10(7).
  • The method effectively distinguishes mutant alleles from abundant wild-type signals, preventing suppression.

Conclusions:

  • Tumorplex™ offers a highly sensitive and specific method for detecting low-abundance clonal mutations in tumor biopsies.
  • The platform's ability to analyze small samples with heterogeneous DNA populations eliminates the need for cell enrichment.
  • Tumorplex™ is a valuable tool for clinical molecular testing, enabling robust mutation detection.