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TUNEL is a method for detecting apoptotic DNA fragmentation, widely used to identify and quantify apoptotic cells, or to detect excessive DNA breakage in individual cells. [3] The assay relies on the use of terminal deoxynucleotidyl transferase (TdT), an enzyme that catalyzes attachment of deoxynucleotides, tagged with a fluorochrome or another ...
EPIC-seq has demonstrated remarkable potential in noninvasive cancer detection, notably in the diagnosis of lung cancer, the leading cause of cancer-related mortality. Using EPIC-seq, researchers have achieved high accuracy in distinguishing between NSCLC patients, DLBCL patients and healthy individuals.
Apoptotic DNA fragmentation is a natural fragmentation that cells perform in apoptosis (programmed cell death). DNA fragmentation is a biochemical hallmark of apoptosis.In dying cells, DNA is cleaved by an endonuclease that fragments the chromatin into nucleosomal units, which are multiples of about 180-bp oligomers and appear as a DNA ladder when run on an agarose gel. [8]
Flow cytometry is widely applied to detect sperm cells abnormality associated with DNA fragmentation [46] in male fertility assays. [47] Also, it is extensively used in research for the detection of DNA damage , [ 48 ] [ 49 ] caspase cleavage and apoptosis . [ 50 ]
Sperm Chromatin Structure Assay (SCSA) is a diagnostic approach that detects sperm abnormality with a large extent of DNA fragmentation. [1] First described by Evenson in 1980, the assay is a flow cytometric test that detects the vulnerability of sperm DNA to acid-induced denaturation DNA in situ. [2]
Fragmented gDNA is incubated with the DNA-RNA structure-specific S9.6 mAb. This step is unique for the DRIP-seq protocol, since it entirely relies on the high specificity and affinity of the S9.6 mAb for DNA-RNA hybrids. The antibody will recognize and bind these regions dispersed across the genome and will be used for immunoprecipitation.
The apoptotic DNA fragmentation is being used as a marker of apoptosis and for identification of apoptotic cells either via the DNA laddering assay, [2] the TUNEL assay, [3] [4] or the by detection of cells with fractional DNA content ("sub G 1 cells") on DNA content frequency histograms e.g. as in the Nicoletti assay.
Molecular diagnostics tool can be used for cancer risk assessment. For example, the BRCA1/2 test by Myriad Genetics assesses women for lifetime risk of breast cancer. [22] Also, some cancers are not always employed with clear symptoms. It is useful to analyze people when they do not show obvious symptoms and thus can detect cancer at early stages.