Submit Article |Resources | Link to us | Sitemap | Search | Language
Back to Home
Home >> Generation of Clones - Generation of Insert >> Nucleic Acid Hybridization Method
Back to Home

Nucleic Acid Hybridization Method - This method was developed by Grunstein and Hogness in 1975 and is used to rapidly determine the colony which contains the sequence from hundreds of clones. In this method, bacterial colonies are transferred onto the nitrocellulose membrane from the agar plate and lysed by using alkaline solution.

When the bacterial cell wall breaks, the proteins and DNA will stick to the membrane, due to the negative charge of membrane. Then the membrane is placed in proteinase K solution to remove the protein bound to the membrane and DNA.

Then the DNA is fixed to the membrane by exposing it to UV rays. After baking, the membrane is exposed to labelled RNA for hybridization. Then RNA-DNA hybrids are monitored by autoradiography.

A colony which gives a positive autoradiography result can then be picked from the master plate. A great advantage of the hybridization method is generality. It does not require expression of the insert sequences and can be applied to any sequence, if suitable probe is available.

 

  

 
 

 
Submit Article |Resources | Link to us | Sitemap | Search | Language
English|Français|Español|Deutsch|Italiano|Portugues|Japanese|Korean|Sim-Chinese| Language