Author: admin
Preparing RNA free of DNA
DNA-free RNA preparations are absolutely required for applications like gene expression analysis using PCR-based methods (e.g., real-time PCR). No RNA…
Preparation of 1M IPTG Solution
Overview: In molecular biology laboratories, IPTG is commonly used to induce the expression of recombinant proteins in E. coli and…
Cell Line : B-16V
Overview Organism Mouse (Mus musculus) Source/Tissue Melanoma Morphology Epithelial-like Cell type Growth mode Adherent, monolayer Doubling time ≈…
Cell Line : A-204
OVERVIEW Organism Human (Homo sapiens), 1 year old, female Source/Tissue Muscle, rhabdomyosarcom Morphology Epithelial-like Cell type Growth mode…
Preparation of Tris-saturated Phenol
Overview: Phenol is a colorless crystalline solid. Phenol gradually turns pink to brownish color due to oxidation upon exposure to…
Preparation of 4% Paraformaldehyde Solution in PBS
A 100 ml of 4% (w/v) paraformaldehyde can be prepared by dissolving 4g of paraformaldehyde in PBS to a final volume of 100 ml.
Detection of DNA in Agarose Gel
♦ After the gel electrophoresis has been completed, the next step is to analyze agarose gel for DNA fragments. Unfortunately,…
Nucleic Acid Staining Dyes for Detection of DNA in Agarose Gel
There are many nucleic acid staining dyes that can be used to visualize DNA embedded in the agarose gel. These…
Preparation of Metaphase Chromosome Spreads from Adherent Cell Line
Overview Preparation of metaphase chromosome spread is an important technique in cytogenetics. Metaphase chromosome spread is used for karyotyping including…
Handling and Aliquoting Frozen Stock of Serum
The serum is commercially supplied in different sizes with 500 ml being the most common size. Since repeated freezing and…
Handling and Aliquoting Frozen Stock of L-glutamine
Ready to use L-glutamine solution (200 mM, sterile-filtered, 100 x concentration) is commercially supplied in different sizes with 100 ml…
Preparing Laminar Flow Hood for Cell Culture Work
Laminar flow hood (Class II) provides a sterile environment for handling cell culture. To make the laminar flow hood ready…
Protocol – Heat Inactivation of Serum
Overview Heat inactivation of serum refers to the process which involves treatment of serum at a higher temperature to inactivate…
Protocol – Feeding Suspension Culture (Complete Replacement of Medium)
Complete replacement of culture medium from suspension culture involves cell harvesting by centrifugation and resuspension of the cell pellet in fresh growth medium. This method is ideally the best way to feed suspension culture and is often used when a. you observe dead or sick cells/suspended particles in culture; b. medium color (phenol red-containing medium) has turned yellow very quickly; c. you can not afford cell loss. Complete replacement of culture medium is ideally the best way to feed suspension culture. Since this procedure takes time and involves an additional centrifugation step to harvest cells, often researchers partially replace the medium that takes just a few minutes. However, partial medium replacement is associated with the loss of cells from the culture.
Protocol – Feeding Suspension Culture (Partial Replacement of Medium)
Replacement of culture medium from suspension culture is a bit tricky job as cells float in the medium. Ideally, cells are harvested from the medium by centrifugation and then resuspended in a fresh culture medium. However, in practice, a part of the culture medium is replaced with a fresh culture medium without harvesting the cells. This procedure is associated with the loss of cells.