Blotting is used in molecular biology to transfer nucleic acids and proteins from gel to a membrane for identification and analysis. Developed in the 1970s, it combines electrophoresis and immunological methods. There are three main types: Southern (DNA), Northern (RNA), and Western (proteins), each allowing detection and measurement of specific molecules.
BLOTTING TECHNIQUE Read More »
Biotechnology, Molecular Microbiology, Techniques in Microbiology Lab
Western blotting technique or protein immunoblot is used to identify specific proteins separated according to their sizes in an electrophoresis experiment. Protein immunoblot unlike other blotting techniques (e.g. southern and northern blotting) utilizes specific antibodies to identify the protein of interest. It is the transfer of proteins from gel to a matrix or nitrocellulose membrane.
WESTERN BLOTTING TECHNIQUE Read More »
Biotechnology, Molecular Microbiology
Northern blotting technique is used to detect specific sequences of ribonucleic acid (RNA). The protocol for performing northern blotting technique is similar to that of southern blotting technique. Northern blotting technique was first described by James Alwine and colleagues in 1977 as a molecular biology technique for identifying specific nucleotide sequence of a piece of
NORTHERN BLOTTING TECHNIQUE Read More »
Biotechnology, Molecular Microbiology
Southern blotting, developed by Sir Edward M. Southern in 1975, is a molecular technique used to detect specific DNA sequences. It involves transferring DNA from a gel to a nitrocellulose membrane, followed by hybridization with radiolabeled probes. This method is pivotal in DNA analysis, forensic science, and paternity testing.
SOUTHERN BLOTTING TECHNIQUE Read More »
Biotechnology, Molecular Microbiology
Microorganisms must meet stringent criteria to be used as probiotics, including being non-pathogenic, non-toxic, and able to survive stomach acid and pancreatic secretions. Probiotics confer health benefits such as enhancing the immune system, producing vitamins, preventing GI infections, and aiding in lactose digestion, potentially treating conditions like IBS and eczema.
Features of probiotics & reported modes of action of probiotics Read More »
Biotechnology, Food Microbiology
A fermenter or bioreactor is simply defined as an apparatus that maintains optimal conditions for the optimum growth of microorganisms used in large-scale fermentation and in the commercial production of economically useful products. Fermenter can also mean fermentation vessel (i.e. an apparatus in which microbes including bacteria, yeasts and moulds causes fermentation to take place).
DESIGN AND OPERATION OF THE FERMENTER Read More »
Biotechnology, Food Microbiology, Industrial Microbiology
STRAIN IMPROVEMENT Strain improvement is defined as the process of improving the production and yielding capacity of a microorganism through certain (deliberate) technological, microbiological, biotechnological, or biochemical process. Microorganisms of industrial importance must possess some certain qualities– which will determine their usage for industrial productions. After their isolation from their natural habitat or environment, the
STRAIN IMPROVEMENT Read More »
Biotechnology, Industrial Microbiology
Melting curve analysis and detection systems Melting curve analysis can only be performed with realtime PCR detection technologies in which the fluorophore remains associated with the amplicon. Amplifications that have used SYBR® Green I or SYBR® GreenER™ dye can be subjected to melting curve analysis. Dual-labeled probe detection systems such as TaqMan® probes are not
Melting curve analysis in Real-time PCR Read More »
Biotechnology, Molecular Microbiology, Techniques in Microbiology Lab
TaqMan® probe signal production Whether an MGB or non-MGB probe is chosen, both follow the same pattern for signal production. In the early PCR cycles, only the low, quenched reporter signal is detected. This early signal, automatically subtracted to zero in the real-time PCR software, is termed “baseline”. If the sample contains a target, eventually
Real-time PCR probes Read More »
Biotechnology, Molecular Microbiology, Techniques in Microbiology Lab
Real-time PCR fluorescence detection systems Real-time fluorescent PCR chemistries Many real-time fluorescent PCR chemistries exist, but the most widely used are 5” nuclease assays such as TaqMan®Assays and SYBR® Green dye–based assays (Figure 1). The 5” nuclease assay is named for the 5” nuclease activityassociated with Taq DNA polymerase (Figure 2). The 5” nuclease domain
Real-time PCR fluorescence detection systems Read More »
Biotechnology, Molecular Microbiology, Techniques in Microbiology Lab