Which of the following steps are involved in recombinant DNA technology?

Options

A. The cells having desired DNA molecule are separated and cultured to increase their number
B. A suitable vector and donor DNA both are treated with similar restriction endonucleases
C. self ligation is increased with alkaline phosphate
D. The host cell cannot be a bacterium

Answer

To answer the question regarding the steps involved in recombinant DNA technology, we can break down the process into six key steps:

### Step-by-Step Solution:

1. **Isolation of Genetic Material**:
- The first step involves isolating the DNA from the organism that contains the desired gene. This is crucial as it provides the genetic material needed for further manipulation.

2. **Restriction Enzyme Digestion**:
- The isolated DNA is then cut into smaller fragments using specific enzymes known as restriction endonucleases. These enzymes recognize specific sequences in the DNA and cleave it, allowing for the desired gene to be extracted.

3. **Using PCR Technique**:
- The Polymerase Chain Reaction (PCR) technique is employed to amplify the specific DNA fragment. This step is essential to produce enough copies of the desired gene for further experimentation and analysis.

4. **Ligation of DNA Molecules**:
- The next step involves ligating the amplified DNA fragment with a vector (a DNA molecule that can carry foreign DNA into a host cell). This is done using the enzyme DNA ligase, which joins the DNA fragments together.

5. **Inserting Recombinant DNA into Host Cells**:
- The recombinant DNA (the vector with the inserted gene) is then introduced into a host organism (such as bacteria). This process is often facilitated by methods like transformation, transfection, or electroporation.

6. **Isolation of Recombinant Cells**:
- Finally, the cells that have successfully taken up the recombinant DNA are isolated and cultured to increase their number. This allows for the study and utilization of the genetically modified organisms.

### Summary of Steps:
1. Isolation of genetic material
2. Restriction enzyme digestion
3. Using PCR technique for amplification
4. Ligation of DNA molecules
5. Inserting recombinant DNA into host cells
6. Isolation of recombinant cells