However, even after transformation, it is essential to confirm the integration of genes, for the success of transgenesis. The incorporation can be confirmed through (i) The use of markers, (ii) Southern blotting, (iii) Northern blotting, and (iv) Polymerase chain reaction. (i) Marker and Reporter Gene: Transformation in eukaryotic system is a very low frequency event. It is, therefore, important that [...]
The following points highlight the seven main stages of recombinant DNA technology. The stages are: 1. Isolation of the Genetic Material (DNA) 2. Cutting of DNA at Specific Locations 3. Isolation of Desired DNA Fragment 4. Amplification of Gene of Interest using PCR 5. Ligation of DNA Fragment into a Vector 6. Insertion of Recombinant DNA into the Host Cell/Organisms [...]
The following points highlight the four main features that are required to facilitate cloning into a vector. The features are: 1. Origin of Replication 2. Selectable Marker 3. Cloning (Recognition) Sites 4. Vectors for Cloning Genes in Plants and Animals. Feature # 1. Origin of Replication (Ori): This is a sequence from where replication starts and any piece of DNA, [...]