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DNA sample is mixed with DNA loading dye (also called sample loading dye) prior to loading onto the wells of agarose gel for electrophoresis. DNA loading dyes contain a high-density reagent and tracking dyes. DNA loading dye makes the DNA sample heavier and coloured, thus helping the DNA sample to sink into the agarose wells without diffusing out and enabling us to monitor the loading process. In addition, tracking dyes in loading dye migrate as a diffuse band to the same direction as DNA which can be seen visually thus allowing us to monitor the progression of electrophoresis. However, tracking dyes sometimes interfere in the analysis of DNA bands by masking it.

10 ml of 6x DNA loading dye containing Bromophenol blue, Xylene cyanol FF, and Ficoll 400 is prepared by dissolving 25 mg bromophenol blue, 25 mg xylene cyanol FF and 1.5 g Ficoll 400 in water to a final volume of 10 ml. The final solution (6x DNA loading dye) will contain 0.25% (w/v) bromophenol blue, 0.25% (w/v) xylene cyanol FF, and 15% (w/v) Ficoll 400.

Overview A DNA sample is mixed with DNA loading dye prior to loading into the wells of agarose gel. Two…

EDTA’s ability to sequester divalent cations from solution without any precipitation makes it a suitable choice to stop an ongoing enzymatic reaction or protect DNA from degradation. Disodium salt of EDTA (EDTA.Na2.2H2O) is most commonly used to prepare 0.5 M EDTA stock solution. A 0.5 M EDTA solution can be prepared by dissolving 186.12 grams of EDTA.Na2.2H2O (Molecular Weight: 372.24) in water to a final volume of 1000 ml.

Overview A serum-free freezing medium is used to preserve cell lines which are maintained in the serum-free growth medium. Serum-free…

Subculturing/passaging can be defined as the preparation of fresh culture by transferring cells from an existing culture. Subculturing is done…

Bacterial contamination is one of the most common cell culture contamination. Poor aseptic culture conditions, including handling, incubator and laminar…

Overview The Trypsin-EDTA method, also referred to as trypsinization, is the most commonly used method for passaging/subculturing of adherent cells….

Suspension culture is passaged by diluting the existing culture. Since cells float in the medium in suspension culture, they are not treated with a trypsin-EDTA solution. To subculture a suspension cell line, a small amount of cell suspension from the existing culture is transferred to a culture dish containing fresh growth medium.

Subculturing (also called passaging) involves transferring a portion of cells from an existing culture to a new culture dish that…

Several methods have been developed for passaging cells. Each method has its own advantages and drawbacks. Always check the cell…

Cryopreservation is an efficient way to preserve cells at ultra-low temperatures (below -135°C) which stops all physiological processes and biological aging. It is a routinely used technique in all cell culture laboratories.

Culturing cells from multicellular organisms especially from animals (mammals, amphibians, insects, birds etc) refers to cell culture or animal cell culture.

Overview The freezing medium is used to preserve cell lines at ultra-low temperatures. This method of preserving cell lines is…

Overview Cryopreservation (Cryo: icy cold or frost; Preservation: Storage) is an efficient way of preserving cell culture at ultra-low temperatures…