What can be more accurate than real time PCR?

“People are different from each other in ways that are fascinating and medically important, and to understand the ways in which variations in our genome give rise to those differences presents an important and interesting set of questions,” says Dr Steve McCarroll, faculty member and principal investigator with the Department of Genetics at Harvard Medical School and the Broad Institute of Harvard and MIT. The McCarroll laboratory studies the biological effects of human genome polymorphism, seeking to define how genome variation influences gene expression and risk of disease. Their work requires them to accurately measure the copy number of a genomic sequence with integer precision in hundreds of people. While real-time PCR and CGH (comparative genomic hybridization) arrays are useful for measuring simple deletions and duplications, these techniques simply weren’t sufficient for obtaining single-molecule resolution of target sequences with extreme precision and accuracy. That’s where Droplet Digital PCR entered the story.

Learn how Dr. McCaroll’s lab, along with other labs across the globe utilize ddPCR in applications such as:

  • Absolute quantification of HIV proviral DNA and human genomic DNA in the same reaction
  • Absolute quantification of HIV viral RNA using two-step reverse transcription-ddPCR
  • Rare event detection of BRAF V600E, EGFR L858R and T790M, JAK-2, and cKIT (cancer mutation analysis and drug resistance mutation)
  • Absolute quantification of miRNA from circulating nucleic acids for biomarker identification
  • High-resolution copy number variation (CNV) analysis of human and mouse DNA’s
  • High-resolution CNV analysis of the MET gene, association study for lung cancer
  • Absolute quantification of next generation sequencing library
  • Rare event detection, mitochondrial mutagenesis
  • Absolute quantification of the BCR-ABL (Philadelphia chromosome) fusion gene and transcript, association with leukemia
  • Absolute quantification and sex determination of fetal DNA in circulating nucleic acid purified from maternal blood

For more information read “Third Generation PCR: A Novel Way to Accurately Measure Copy Number Variation in the Human Genome.”

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