Gel Electrophoresis

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[edit] Gel Electrophoresis

Gel Electrophoresis is a technique used for the separation of deoxyribonucleic acid, ribonucleic acid, protein molecules using a electric current applied to a gel matrix. It is usually performed for analytical purpose but may be used as the preparative techniques prior to use of other methods such as mass spectrometry, RFLP, PCR, cloning, DNA sequencing.

[edit] Details

Agarose, acrylamide(polyacrylamide gel, SDS-polyacrylamide gel),Buffer(TBF) are requirement.

Buffer provides ions in the solution to ensure electricity conductivity power supply gel chamber,cellulose acetate.

Remember that DNA is the organic molecule, and is negatively charged. When electric field is applied to it, particles migrate towards positive electrode. An agrose gel is used to slow the movement of DNA and separate it by size.

How fast DNA will migrate depends upon the strength of electric field, buffer, density of agarose gel and size of the DNA.Smaller the size of DNA, faster it will move.Within an agarose gel, linear DNA migrate inversely proportional to the log10 of their molecular weight.

[edit] 2-D Electrophoresis

It is form of gel electrophoresis commonly used to analyze the protein.Mixtures of protein are separated by two properties in two dimension on 2-D gel.

Two separation occurs

1.Separation of protein in first direction on the basis of charge by isoelectric focussing.

2.The resultant gel strip is applied to an SDS(Sodium Dodecyl sulphate) polyacrylamide ge and the proteins are separated into bands by mass.This separation is in 2nd Dimension by size.To see the bands one must stain the gel with ethidium bromide.