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Home >> Blotting Techniques >> Plaque OR Colony Blotting Techniques
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Plaque OR Colony Blotting Techniques - This method was first developed by Granstiens and Hogness (1975). This method is used to identify which colony of bacteria contains the DNA of interest among thousands. In this procedure, the bacterial colonies to be screened are transferred onto nitrocellulose or nylon membrane by using replica plating.

Due to the negative charge of the cell surface, some cells bind to the nitrocellulose membrane. Then the membrane is placed in a solution of 0.5 N NaOH to break the cell surface, convert dsDNA to ssDNA and to bind DNA to the membrane. Later, the membrane is transferred to a solution containing protease solution after neutralizing with neutralization solution.

The DNA is fixed tightly to membrane by either W cross linking or oven baking. This membrane is used for hybridization with a probe and analysed by using autoradiography or biotin method for positive hybridization. A colony whose DNA print (as replica plating provides a replica print master plate colony on the membrane) gives a positive hybridization can be picked from the master plate.

Plaque blotting is similar to colony blotting, the only difference is that instead of bacterial colony, a plaque is transferred onto the membrane. Benton and Davis developed this method in 1977. The greatest advantage of this method is that several identical DNA prints can be easily made from a single master plate containing bacteria/plaques which are to be made.

 
 

 

 
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