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Preparation of 6X DNA Loading Dye (Bromophenol blue, Xylene Cyanol FF, Ficoll 400)

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.

Preparation of 0.5 M EDTA Solution from Disodium EthyleneDiamineTetraacetate Dihydrate (EDTA.Na2.2H2O)

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.

Protocol – Cryopreservation of Adherent Cells Growing in Serum-free Medium

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

Subculturing adherent cells using trypsin-EDTA

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

Bacterial Contamination in Cell Culture

Bacterial Contamination in Cell Culture

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

Protocol – Subculturing Adherent Cells Growing in Serum-containing Growth Medium using Trypsin-EDTA

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

Protocol – Passaging/Subculturing Suspension Culture

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.

Passaging/subculturing cells

Cell culture is not static. Cells acquire changes when maintained for a long time in culture (Masters & Stacey, 2007)….

Passaging/Subculturing Methods for Cells

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

Cryopreservation of Cells

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.

Cell culture overview

Cell Culture

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

Preparation of Freezing Medium Containing DMSO and Serum

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

Protocol – Cryopreservation of Adherent Cell Culture

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