Author: admin
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.
Preparation of 10 M Sodium Hydroxide (NaOH) Solution
Here we have described a procedure to prepare 10 M (or 10 N) sodium hydroxide (NaOH) solution. A 10 M NaOH solution can be prepared by dissolving 40 g of NaOH (Molecular Weight: 40.00) in water to a final volume of 100 ml. Since dissolution of NaOH in water is a highly exothermic reaction, extreme care must be taken while preparing the solution.
Protocol – Feeding Adherent Culture
Feeding adherent culture involves replacing the old consumed medium with the fresh growth medium. The purpose is to supply fresh…
Preparation of Stock Solution of L-Glutamine (200 mM)
Overview L-Glutamine (C5H10N2O3; CAS number: 56-85-9) is an important component of the cell culture medium. It is an alternative energy…
Feeding Cells
Feeding is an essential step to maintain cells in a healthy state in culture. Feeding of a culture is a…
Protocol – Thawing and Revival of Cryopreserved Cells
This protocol describes the revival process of cryopreserved adherent cells. The revival process involves rapid thawing of cryopreserved cells followed by removal of freezing medium which contains cryoprotectant, in this case, DMSO. The freezing medium can be removed immediately just after thawing the cells by centrifugation. Alternatively, cells are allowed to adhere to the culture dish followed by washing and addition of fresh growth medium.
Cell Line : A7r5
Overview Organism Rat (Rattus norvegicus), Strain DB1X Source/Tissue Embryo, Thoracic aorta, Medial and intimal layers Morphology Fibroblast-like Cell type…
Preparation of lysozyme stock solution (10 mg/ml)
Lysozyme is an enzyme used for the extraction of proteins and nucleic acid from bacteria. It improves the extraction process…
Protocol – Preparing Liquid Culture of E. coli for Plasmid Miniprep
Overview:
Small-scale plasmid isolation, called plasmid miniprep, requires a small amount of bacterial culture. Normally, a 2 – 5 ml culture provides sufficient cells that can be processed for plasmid isolation using the miniprep protocol. Culture can be prepared by inoculating 3 ml antibiotic-containing growth medium with a single colony and growing it overnight at 37C with shaking.
Here we have taken an example of preparing liquid culture from a colony of E. coli DH5α, transformed with the pEGFP plasmid. The pEGFP plasmid contains a kanamycin resistance gene, therefore, requires kanamycin for the selection of plasmid-containing bacteria. If your plasmid carries another antibiotic-resistant gene, add the respective antibiotic to the culture medium.
Preparation of Glucose-containing Resuspension Buffer for Plasmid Isolation by Alkaline Lysis Method
Overview: Resuspension buffer is used to resuspend the harvested bacterial cells from the culture during plasmid isolation by the alkaline…
Preparation of Neutralization Solution (Solution III) for Plasmid Isolation by Alkaline Lysis Method
Overview The neutralization solution (solution III) is used for the isolation of plasmid DNA by the alkaline lysis method. The…
Preparation of Lysis Solution (Solution II) for Isolation of Plasmid by Alkaline Lysis Method
Overview Lysis solution (solution II) is used for the isolation of plasmid DNA by the alkaline lysis method. The plasmid-containing…
Suppliers – 0.5M EDTA solution
Note: We have listed some of the suppliers. You must visit the supplier’s homepage to order and to get more…
Preparation of 0.5 M EDTA Stock Solution from Anhydrous EDTA Free Acid
EDTA (EthyleneDiamineTetraAcetic acid), a polyamino carboxylic acid, is extensively used in molecular biology experiments as a chelating agent. It sequesters…
Preparation of 1 M Tris.Cl Buffer
Overview Tris is also known as THAM, Tris base, Trizma, Tris(hydroxymethyl)aminomethane. Tris is one of the most frequently used buffers…