1 How much agarose would you need to make 46mL of a 09 agaro

1.) How much agarose would you need to make 46mL of a 0.9% agarose gel? Show all work.

2.) What is the point of running DNA out in a gel?

Solution

1. ) 0.9 % of agrose gel = 0.9 gm in 100ml of TAE (Tris acetate EDTA)

So for 46 ml of agrose gel the quantity of agrose needed would be:

(0.9/100)*46 = 0.414 gm

Hence 0.414 gm of agrose would be required.

2.) Electrophoresis is described as a technique for separating molecules (DNA, RNA ,Proteins etc.) in a mixture when an electric field is applied. The molecules migrate under the influence of applied electric field at a speed which is determined by their charge:mass ratio. For example, if two molecules have the same mass and shape, the one with the greater net charge will move faster toward an electrode.

Gel is a crosslinked polymer whose porosity and composition is chosen based on the specific weight and composition of the target to be analyzed. Gels used in gel electrophoresis act as an anticonvective medium or sieving medium during migration of molecules. They suppress the thermal convection caused by application of the electric field, and can also act as a sieving medium, retarding the passage of molecules. Random diffusion of the molecules is restricted by gel matrix which results in migration of molecules of the same length together as a band whose width equals that of the well into which the original DNA mixture was placed at the start of the electrophoretic run. Further agrose gel which is used for DNA separation is easy to cast and has relatively fewer charged groups which helps in greater range of DNA separation. It also has good gel strength and large pore size making it suitable aniticonvection agent.