KARYOTYPING LAB

J774 cell line used in this experiment is from a BALB/c/NIH mouse with reticulum cell sarcoma. the cells are known for their ability to perform antibody-dependent phagocytosis, making them helpful tools for investigating immune responses to various antigens. Colcemid was used in chromosome analysis during karyotyping. This treatment with Colcemid prevents spindle formation during mitosis, causing metaphase arrest. Metaphase is the optimum phase of mitosis for microscopically visualizing the chromosomes. Giemsa is a visible light dye that binds to DNA through intercalation and thus s used for chromosome staining the chromatin is stained red and cytoplasm blue-banding or Giemsa banding is a technique used in cytogenetics to produce a visible Karyotype by staining condensed chromosomes. 

Typically, karyotyping is performed using peripheral blood leucocytes (isolated from a heparinized blood sample), which are initially cultured, before evaluation by high-resolution G-banding (Giemsa staining) of at least 20 metaphase nuclei.