DBT Builder National Facility

Epigenetics and Genome Editing

Droplet Digital Polymerase Chain Reaction (ddPCR):

The Droplet Digital Polymerase Chain Reaction (ddPCR) instrument offers a significant advancement over traditional PCR methods by providing highly precise and sensitive quantification of nucleic acids. Unlike conventional PCR, which relies on exponential amplification and is prone to variability, ddPCR partitions the sample into thousands of individual droplets, allowing for absolute quantification without the need for standard curves. This droplet-based system enables the detection of rare genetic mutations, copy number variations, and low-abundance targets with greater accuracy.

Additionally, ddPCR minimizes the effects of PCR inhibitors and enhances reproducibility, making it an ideal choice for applications in cancer research, gene expression analysis, and pathogen detection. By choosing ddPCR, researchers can achieve more reliable results, paving the way for groundbreaking discoveries in molecular biology.

ddPCR is a versatile tool with a wide range of applications in molecular biology, including:

• Quantifying circulating miRNA : Enables precise measurement of miRNA levels in biofluids, important for biomarker discovery and disease monitoring
• Measuring Double-Strand Breaks (DSBs) at Cas9 cut sites: Offers accurate detection and quantification of DSBs, enhancing the evaluation of CRISPR/Cas9 gene editing efficiency.
• Detecting KRAS mutations in exosomal DNA from pancreatic ductal adenocarcinoma (PDAC) patients: Facilitates early detection of cancer-related mutations in a minimally invasive manner.
• Detecting the presence of trisomy 21: Provides highly sensitive and reliable prenatal screening for Down syndrome.
• Confirming low-frequency mosaic CNVs in parental fibroblasts: Assists in the detection of rare copy number variations, crucial for understanding genetic mosaicism.
• Detecting paternal CFTR mutations in Cell-Free Fetal DNA (cffDNA) from maternal plasma: Enables non-invasive prenatal testing for cystic fibrosis
• Detecting both common and rare deletions in thalassemia: Allows for precise identification of deletions, aiding in the diagnosis and management of thalassemia.

Fluorescence-activated cell sorting (FACS):

Fluorescence-activated cell sorting (FACS) is a powerful and versatile instrument widely used in biomedical research for cell analysis and sorting. FACS allows for the precise identification and isolation of specific cell populations based on fluorescent labeling. Unlike traditional cell sorting methods, FACS enables high-throughput sorting of individual cells with exceptional accuracy, even within heterogeneous mixtures. This technology is particularly valuable for applications such as:

• Immunophenotyping: : Characterizing and sorting immune cell subsets based on surface markers.
• Stem Cell Research : Isolating stem cells for regenerative medicine or research.
• Cancer Research : Identifying and sorting cancer stem cells, rare tumor cells, or cells expressing specific mutations.
• Single-Cell Analysis: Providing a detailed understanding of cellular heterogeneity by sorting cells for downstream genomic or proteomic analysis.

The precision, speed, and ability to sort cells based on multiple parameters simultaneously make FACS an essential tool for modern cell biology and clinical diagnostics.