Student need to know How Genomic DNA extraction works?

While there may be a number of ways by which you can successfully isolate your genomic DNA from your sample, your final choice of which genomic DNA extraction protocol to use will ultimately depend on several factors which includes the following:

• the molecular weight of the DNA of interest;
• quantity and purity required to facilitate downstream applications;
• ease or complexity of your chosen method; 
• time requirements; and 
• budgetary constraints.

Genomic DNA can be separated from all other cellular components by simply following these three basic steps:

Disruption and cell lysis. In extracting your genomic DNA from the sample, you need to break down the cell walls that protect the DNA by using enzymes such as lysozyme and proteinase K or by using physical methods such as manual grinding (mortar and pestle method), freeze-thaw technique, sonication, liquid homogenization and/or mechanical disruption with the use of a Waring blender or a polytron. You can also use bead beating (using 0.1 mm glass beads or 0.15 mm garnet beads) to release your genomic DNA from your cell lysate.

Understanding What assay development accessories should be used for protein binding?

As mentioned in our earlier post, there are several kinds of activated plates that can help you get the results you need to complete your research. Now, since we have already covered biotin-binding plates, we will now take a closer look at some of the most commonly used protein/peptide binding plates.

Protein/Peptide Binding Plates
Plates designed for protein/peptide binding can either be coated with nickel (for binding 6X histidine tagged proteins and peptides), glutathione (for GST tagged proteins and peptides), amine (for binding primary amines of peptides and proteins) and/or sulfhydryl (for free sulfhydryls of peptides and proteins). Most of these plates are supplied as clear, white and black and can ideally be used for colorimetric, chemiluminescent and fluorescent detection methods, respectively.

Nickel coated plates directly isolate polyhistidine-tagged proteins and peptides while glutathione coated plates isolate glutathione S-transferase (GST) tagged proteins from bacterial lysates for ELISA-based protocols. Detergents used to lyse cells do not inhibit protein binding to activated plates as they usually do with plain polystyrene plates. In addition, these plates are ready to use and are pre-blocked to reduce non-specific binding.

Amine coated plates are maleic anhydride activated plates that rapidly bind primary amines of peptides and proteins to form amide bonds that are stable under neutral and basic conditions (pH≥7). Under acidic conditions, however, the bonds will be hydrolyzed releasing the peptide/ligand. Taking this into consideration, binding of peptide/ligand to plates should be performed between pH 8 to 9 and the binding assays or ELISA should be performed at pH conditions ≥7.

Sulfhydryl-binding plates are maleimide activated plates that react with free sulfhydryls to form stable thioether bonds between pH 6.5 and 7.5. These are ideally used in immobilizing and binding sulfhydryl-containing molecules, especially those that are difficult to coat onto polystyrene plates (e.g. protein molecules with free sulfhydryl group and peptides that contain a terminal cysteine). Both amine-binding and sulfhydryl-binding plates are pre-blocked to reduce non-specific binding.

Amazing Harbour Bridge in Sydney

The Sydney Harbour Bridge is one amongst Australia's most documented and photographed landmarks. it's the world's largest (but not the longest) steel arch bridge with the highest of the bridge standing 134 metres higher than the harbour. it's lovingly celebrated by the locals because the 'Coathanger' attributable to its arch-based style.

A history of the Sydney Harbour Bridge

It was as early as 1815 that Francis greenbelt projected building a bridge from the northern to the southern shore of the harbour.

It took it slow for this to become a reality with style submissions invited in 1900. All the submissions were thought-about unsuitable then the momentum for the bridge crossing stopped.

However, when the primary war additional serious plans were created, with a general style for the Sydney Harbour Bridge ready by Dr J J C Bradfield and officers of the authority Department of structure. The New South Wales Government then invited worldwide tenders for the development of the Bridge in 1922 and therefore the contract was let to English firm Dorman Long and Co of Middlesbrough.

The Sydney Harbour Bridge construction started in one924 and took 1,400 men eight years to create at a value of four.2 million. Six million hand driven rivets and fifty three,000 tonnes of steel were utilized in its construction. It currently carries eight traffic lanes and 2 rail lines, one in every direction, however at the time of its construction the 2 japanese lanes were tram tracks. They were regenerate to road traffic once Sydney closed down its tram system within the Fifties.