How does 3H-thymidine work?

How does 3H-thymidine work?

3H-Thymidine is a radioactive version of the Thymine DNA base (thymine + the sugar backbone = thymidine). When cells are incubated with thymidine, they use the radiolabeled thymidine to synthesize DNA and incorporate it into their DNA backbone. Just incubate cells in the presence of BrdU.

What is 3H TDR?

Autoradiography with 3H-Thymidine (3H-TdR) The pyrimidine nucleoside thymidine is a specific precursor of DNA. Injected during the S-phase of the cell cycle, it is incorporated into the cell nucleus as an almost exclusively stable, not exchangable compound.

Who used thymidine radioisotope in DNA replication?

This technology showed that, e.g., thymidine is a precursor for DNA, but not for RNA [9]. In 1957, Taylor and colleagues developed a method where they used H3-thymidine for the labeling of replicated DNA in the bean root, and autoradiography for its detection.

What is the difference between thymidine and thymine?

The key difference between thymine and thymidine is that thymine is a nucleobase, whereas thymidine is a nucleoside. Moreover, thymine is a single planar molecule while thymidine is a combination of two molecules; ribose sugar and thymine.

Is BrdU radioactive?

Because BrdU can replace thymidine during DNA replication, it can cause mutations, and its use is therefore potentially a health hazard. However, because it is neither radioactive nor myelotoxic at labeling concentrations, it is widely preferred for in vivo studies of cancer cell proliferation.

What is BrdU staining?

BrdU (Bromodeoxyuridine / 5-bromo-2′-deoxyuridine) is an analog of the nucleoside thymidine used in the BrdU assay to identify proliferating cells. BrdU antibodies can be used in conjunction with cell type markers such as doublecortin, and NeuN to identify proliferating cells and newly differentiated neurons.

What is tritiated thymidine?

(trĭt′ē-āt″ĭd thī′mĭ-dēn″, trĭsh′) 3. H-Tdr; a radioactively labeled nucleoside used to measure T lymphocyte proliferation in vitro. Thymidine is essential for DNA synthesis; thus the amount of 3H-Tdr taken up is a general measure of the number of new lymphocytes produced.

What is the difference between thymine and thymidine?

Is thymine A nucleoside?

Nucleosides are the structural subunit of nucleic acids such as DNA and RNA. A nucleoside, composed of a nucleobase, is either a pyrimidine (cytosine, thymine or uracil) or a purine (adenine or guanine), a five carbon sugar which is either ribose or deoxyribose.

Is thymine an amino acid?

The genetic code is the sequence of nucleotide bases in nucleic acids (DNA and RNA) that code for amino acid chains in proteins. DNA consists of the four nucleotide bases: adenine (A), guanine (G), cytosine (C) and thymine (T). Amino acids are linked together to form proteins.

How is 3 H thymidine used to monitor DNA synthesis?

Metabolic incorporation of 3 H-thymidine into cellular DNA is a widely used protocol to monitor rates of DNA synthesis and cell proliferation. However, this radiochemical has also been reported to induce cell-cycle arrest and apoptosis in addition to DNA damage.

Where do I get 3 H-thymidine media?

Uniformly 13 C- and 15 N-enriched thymidine [U- 13 C, 15 N-TdR] was obtained from Martek Biosciences (Columbia, MD), and 3 H-TdR (80 Ci/mmol) was purchased from ICN Radiochemicals (Irvine, CA). Unlabeled thymidine came from Sigma (St. Louis, MO). Media and buffers were obtained from Fisher Scientific (Pittsburgh, PA).

What is the purpose of the thymidine incorporation assay?

Assay Description. The thymidine incorporation assay, the most common assay, utilizes a strategy wherein a radioactive nucleoside, 3H-thymidine, is incorporated into new strands of chromosomal DNA during mitotic cell division. A scintillation beta-counter is used to measure the radioactivity in DNA recovered from the cells in order to determine

How is the decay of tritium used in the laboratory?

The unusually low energy released in the tritium beta decay makes the decay (along with that of rhenium-187) appropriate for absolute neutrino mass measurements in the laboratory (the most recent experiment being KATRIN ).