Gene cloning problem

  • #1
1,007
76

Homework Statement



Molecular cloning experiments are carried out to generate multiple copies of a gene of interest. In one such experiment carried out at a student lab, the gene coding for insulin is multiplied in a bacterial cell. In the screening step, the students realized that they have committed a mistake and they observe many transformed and non-transformed bacterial colonies. Explain this situation with appropriate reasoning.

Homework Equations


NA

The Attempt at a Solution


After the restriction enzymes are used, is it that some plastid is not able to recombine with particular gene and the same plastid combine?
 

Answers and Replies

  • #2
Ygggdrasil
Science Advisor
Insights Author
Gold Member
3,337
3,607
After the restriction enzymes are used, is it that some plastid is not able to recombine with particular gene and the same plastid combine?

There are a number of explanations I can think of, but yours seems reasonable. I would not use the word "recombine," however, as it has a specific definition in biology. Rather, the correct term for cloning would be ligation. It is worth considering which step in the cloning procedure usually prevents recircularization of the plasmid during the ligation step.
 
  • #3
1,007
76
There are a number of explanations I can think of, but yours seems reasonable. I would not use the word "recombine," however, as it has a specific definition in biology. Rather, the correct term for cloning would be ligation. It is worth considering which step in the cloning procedure usually prevents recircularization of the plasmid during the ligation step.
But what mistakes the student have done? It will happen naturally always that some will transform and some not.
 
  • #4
Ygggdrasil
Science Advisor
Insights Author
Gold Member
3,337
3,607
What step in the cloning procedure normally prevents recircularization of the plasmid during the ligation step? That is the step that was likely omitted if your hypothesis is true.
 
  • #5
1,007
76
What step in the cloning procedure normally prevents recircularization of the plasmid during the ligation step? That is the step that was likely omitted if your hypothesis is true.
I don't know that step. I searched ligation in wikipedia but was not able to get the step.
 

Related Threads on Gene cloning problem

  • Last Post
Replies
2
Views
3K
  • Last Post
Replies
1
Views
1K
  • Last Post
Replies
1
Views
4K
  • Last Post
Replies
1
Views
1K
  • Last Post
Replies
1
Views
4K
Replies
1
Views
840
Replies
15
Views
2K
  • Last Post
Replies
2
Views
8K
Replies
3
Views
1K
Top