BaselineThe baseline of the real-time PCR reaction refers to the signal level during the initial cycles of PCR, usually cycles 3 to 15, in which there is little change in fluorescent signal. The low-level signal of the baseline can be equated to the background or the “noise” of the reaction (Figure 1). The baseline in […]
Real-time PCR analysis technology Read More »
Biotechnology, Molecular Microbiology, Techniques in Microbiology Lab
Good primer design is one of the most important parameters in real-time PCR. This is why many researchers choose to purchase TaqMan® Assay products—primers and probes for real-time PCR designed using a proven algorithm and trusted by scientists around the world. If you choose to design your own real-time PCR primers, keep in mind that
Real-time PCR primer design Read More »
Biotechnology, Molecular Microbiology, Techniques in Microbiology Lab
DNA polymerasePCR performance is often related to the thermostable DNA polymerase, so enzyme selection is critical to success. One of the main factors affecting PCR specificity is the fact that Taq DNA polymerase has residual activity at low temperatures. Primers can anneal nonspecifically to DNA during reaction setup, allowing the polymerase to synthesize nonspecific product.
REAL-TIME PCR COMPONENTS Read More »
Molecular Microbiology, Techniques in Microbiology Lab
Real-time PCR is a variation of the standard PCR technique that is commonly used to quantify DNA or RNA in a sample. Using sequence-specific primers, the number of copies of a particular DNA or RNA sequence can be determined. By measuring the amount of amplified product at each stage during the PCR cycle, quantification is
STEPS INVOLVED IN PERFORMING REAL-TIME PCR Read More »
Molecular Microbiology, Techniques in Microbiology Lab
The polymerase chain reaction (PCR) is one of the most powerful technologies in molecular biology. Using PCR, specific sequences within a DNA or complementary DNA (cDNA) template can be copied, or “amplified”, many thousand- to a million-fold using sequence-specific oligonucleotides (or primers), heat-stable DNA polymerase, and thermal cycling. In traditional (endpoint) PCR, detection and quantification
INTRODUCTION TO REAL TIME POLYMERASE CHAIN REACTION (RT PCR) Read More »
Biotechnology, Molecular Microbiology, Techniques in Microbiology LabPrimers are short stretches of DNA that target unique sequences of a DNA molecule and help identify a unique part of a genome or a gene to be copied further. They are usually 18 to 25 nucleotides long. Primers can be synthesized and used for your PCR (polymerase chain reaction) and other molecular biology experiment
HOW TO DESIGN PRIMERS FOR YOUR PCR EXPERIMENT Read More »
Biotechnology, Microbe Lab, Molecular Microbiology, Techniques in Microbiology Lab
There are different types of steps involved in the transformation of a bacterial cell in gene cloning techniques. Bacterial cells are transformed when they are infused with exogenous DNA from another organism, and this makes them competent enough to express the gene product (or protein) of the gene they are carrying. Usually, the exogenous DNA
STEPS INVOLVED IN TRANSFORMING BACTERIAL CELLS Read More »
Biotechnology, Molecular Microbiology
Cloning is a molecular biology technique which is used to make millions of copies of a particular gene of interest or a piece of DNA molecule. It generally involves taking a piece of DNA molecule (containing specific gene sequence of interest) from a donor organism where it naturally exists and putting same into a cloning
Mutagenesis is the careful alteration of the genetic information of an organism’s genome in such a way that it will result in a mutation. It is the creation of genetic change in the DNA of an organism and, which will result in the production of a population of cells with an entirely new genetic make-up.
Mutation is a change in the genetic make-up (or DNA) of an organism.It is an inheritable alteration in the nucleotide sequence (or genome base sequence) of a living organism. Mutation can occur in both plant and animal cells, and it can be spontaneous (i.e. natural), prompted (or artificial), or adaptive in nature. When mutation occurs,
