Question

What are the 7 steps of design for a molecular cloning experiment (in order)?

group of answer choices
1. amplifying gene of interest and electrophoresis
2. cleaving dna
3. ligation
4. transformation
5. screening
6. dna purification
7. sequencing

Answer 1

The 7 steps of design for a molecular cloning experiment are: amplifying gene of interest and electrophoresis, cleaving DNA, ligation, transformation, screening, DNA purification, and sequencing.

Step-by-step explanation:

1. Amplifying gene of interest and electrophoresis: The gene of interest is amplified using the polymerase chain reaction (PCR), and the resulting DNA fragments are separated using gel electrophoresis.
   
2. Cleaving DNA: The amplified gene is digested with restriction enzymes, which cut the DNA at specific recognition sites.
   
3. Ligation: The digested gene is ligated into a cloning vector, such as a plasmid, using the enzyme DNA ligase.
   
4. Transformation: The recombinant vector is transformed into host cells, such as bacteria, using a method such as heat shock or electroporation.
   
5. Screening: The transformed cells are screened to identify those containing the desired gene. This can be done by selecting for the presence of a selectable marker or by using a specific probe for the gene of interest.
   
6. DNA purification: The selected transformants are grown and the plasmid DNA containing the gene of interest is purified.
   
7. Sequencing: The purified DNA can be sequenced to confirm the presence and sequence of the gene of interest.