Key Steps of Molecular Cloning

Animation demonstrating the concept and key steps of molecular cloning.
Narration text is on our website: http://www.cloningstrategies.com/animations.html

Category:

Education

Tags:

• Molecular cloning
• DNA
• vector
• plasmid
• insert
• restriction
• restriction enzyme
• ligation
• transformation
• E.coli

[ntlema2]

couple of questions:

How do you determine which restriction enzymes to use?

What liquid medium was used in the end to extract the cloned DNA?

[CloningStrategies]

@ntlema2 Great questions!

Which enzymes to use: This is what you decide as a scientist when you are planning your cloning experiment. You look at the sequence of the plasmid and see what restrictions sites are available, then you integrate those sites into the primers you use to amplify the DNA fragment. Check "Simply Cloning - Chapter 1 - Planning" video.

To extract cloned DNA: It is more than just a liquid medium. Check "Chapter 8 - Plasmid Miniprep"

[powerzoner]

@CloningStrategies why are two different REs used?

[CloningStrategies]

@powerzoner If you stop the video at 2:58 min you will see that overhangs from different restriction enzymes don't match each other. So plasmid vector can't close on its own, only with the insert.

There are all kinds of exceptions, but using two REs is a simple way of preventing vector from closing onto itself.

[feb17aaru]

This is such a helpful video!THANK YOU SO MUCH!

[CloningStrategies]

@feb17aaru

- Hey, you made my day :-)

[biochemisery]

I really like this animation because it makes everything simple. I was rather confused before that.

[shoppittsburghnow]

great vid you have here

[staranjela]

really informative and interesting

[alexasmithy]

very interesting video thanks

[eckostar61]

is there any video on expession DNA cloning?? please need help with it

[pawangurway]

simple... usefull... n effective !!! aWESOME !!!!

[powerzoner]

@CloningStrategies Oh I see! Thanks man, great vid.