Buffers play a critical role in chemistry, biology, and medicine because they maintain a stable pH in solutions despite the addition of small amounts of acids or bases. Among the wide variety of buffer systems available, sodium acetate buffer is one of the most widely used …
Introduction of Salkowski reagent Salkowski reagent is a well-established chemical solution used primarily for the qualitative and semi-quantitative detection of indole compounds, especially indole-3-acetic acid (IAA), the most common naturally occurring plant hormone in the auxin group. Named after the German chemist Ernst Leopold Salkowski, the …
The extraction of DNA from agarose gel has many downstream applications in molecular biology research work and biomedical research. For instance, we often need to sequence or clone the DNA fragments obtained either by PCR or restriction digestion. Therefore, it is necessary to have a pure …
Electroelution is one of the numerous methods available for DNA extraction from agarose gel. Here, I have described some of the specialized electroelution apparatus. These can easily be used to isolate DNA from the agarose gel. Though these apparatuses may be expensive to buy they offer …
Agarose gel extraction can be made easy by using commercially available gel extraction kits or advanced electrophoretic apparatuses that allow the visualization of DNA bands during the electrophoresis. Conventional methods include dialysis of the sliced gel containing DNA by applying an electric current while some other methods include spin columns and Sephadex …
Here, I have described different types of commercially available gel extraction kits for gel excision. These are listed below in a table summarizing the most important comparative information about different kits. These commercially available gel extraction kits can easily be utilized for DNA gel excision. The …
Here, I have described some of the recently developed portable agarose gel electrophoresis systems with the built-in safe imager. The built-in imager or illuminator helps us visualize the DNA migration in real time while doing electrophoresis. These setups have been designed in such a way that …
Urea and formamide are used as denaturants in Denaturing Gradient Gel Electrophoresis (DGGE). In DGGE, urea, and formamide denature the DNA fragments by disrupting the Hydrogen bonding between the nitrogenous base pairs. As shown in the figure above, urea and formamide form a Hydrogen bond with nitrogenous bases …
What is SDS PAGE? SDS PAGE or Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis is an analytical tool widely used to analyze protein mixtures qualitatively as well as quantitatively. However, we can also use it to determine the molecular weight of an unknown protein using the SDS …
What are APS and TEMED? Before jumping into the uses of the APS and TEMED in SDS PAGE, let’s know more about the individual components. APS is the ammonium persulfate while TEMED is the N, N, N’, N’-tetramethyl ethylene diamine. In the same manner, SDS is …
