live-dead-assay-bacteria-clostridium-sporogenes

TUNEL assay is the cell death detection method where the biochemical marker of apoptosis is DNA fragmentation. The assay involves the microscopical detection of generated DNA fragments with free 3'-hydroxyl residues. in apoptotic cells using enzyme terminal deoxynucleotidyl transferase (TdT) which adds biotinylated nucleotides at the site of DNA breaks. Major challenges of this method involve proper access of the enzyme which could be hampered by poor permeabilization and/or excessive fixation with cross-linking fixative (common with archival tissue). This issue can be resolved by optimizing the incubation time with Proteinase K or CytoninTM.

Bacterial culture is a process of letting bacteria multiply in a controlled fashion (temperature, humidity, oxygen content or shaking), in a predetermined culture medium (antibiotic resistance to obtain homogenous clones). It is an important step, especially during cloning, as a single cell can be grown homogeneously (on semi-solid or in liquid conditions) to obtain colonies. As mentioned, bacteria can be cultured in broth cultures (Luria broth or LB) or Petri dishes (Agar plates). A specific antibiotic can be added to the broth or agar plates in order to grow bacteria which have the gene insert conferring its resistance to that antibiotic. Following points are necessary to consider for optimal growth conditions: 1. In general, most bacteria grow well at 37C, but there are some strains which require growth temperatures between 25-30C. 2. It is ideal in broth cultures to fill the flask to ⅓ or less of the total flask volume for optimal aerobic growth. 3. Shaking speeds between 140-180 rpm are appropriate to ensure aeration and that the cells are surrounded by fresh media, and do not settle.

Get tips on using DeadEnd™ Colorimetric TUNEL System to perform TUNEL assay cell type - Mouse liver tissue

Get tips on using Host Cell Residual DNA contamination LANCE Ultra TR-FRET Detection Kit, 500 Assay Points to perform Cell Culture Contamination Detection Kit Bacteria

Get tips on using TRIzol Reagent to perform RNA isolation / purification Bacteria - Gram positive Clostridum botulinum

Get tips on using RNeasy Mini Kit to perform RNA isolation / purification Bacteria - Gram positive Clostridum botulinum

Get tips on using ReliaPrep™ RNA Miniprep Systems to perform RNA isolation / purification Bacteria - Gram positive Clostridum botulinum