Restriction Enzyme Analysis is something we use everyday.
And it’s almost something that has become so routine that lots of times we don’t think very much about it.
But in fact, it’s really a very cool process.
So, my laboratory is interested in cancer and cancer biology.
And we are most interested in trying to understand the genes whose mutation
give rise to cancer and trying to think of ways to exploit that to selectively kill cancer cells.
Restriction enzymes cut specific sequences of DNA,
and that sequence can be different between different restriction enzymes.
And then the trick is to try to separate the DNA that has been cleaved.
And so one uses a technique called Gel Electrophoresis
because of course, DNA is charged, it will move through an electric current.
And because the gel is porous, some molecules move through it faster than others.
So, if they’re smaller, they go through it faster and if they’re bigger, they go through it slower.
And as a consequence, you can separate them by size.
For engineering recombinant DNA, one can identify the different fragments because they have been separated by size.
We can then move that into, next to other pieces of DNA, and using a different DNA ligation
begin to splice different DNA fragments together.
And that enables us, as scientists, to control when a gene is on and off
as opposed to the way it would naturally be controlled.
And we can use that to learn things about many cellular processes including cancer.
probably every lab here at the Sloan-Kettering to one degree or another uses Restriction Enzymes in their work.
There’s a lot of techniques in Molecular Biology that by being able to digest DNA at specific sites,
one can use that for various applications.
What are you loading into your gel right now?
Right now we are loading our negative control into the gel.
What's the negative control?
The negative control would be water.
Deionized water.
What else have you loaded so far?
So far we've loaded our molecular weight marker, EcoR1, and HindIII, which are all enzymes.
What do you have to be careful of when you load?
When you load the gels, you have to be careful that you don't go in too far and puncture the gel,
and you also want to make sure that you don't miss the well and get the sample everywhere, floating through the buffer.
We can figure out where a specific enzyme cuts.
So each enzyme has a recognition sequence that where it cuts DNA.
We have a mystery enzyme, and we could figure out what enzyme that is based on where HindIII and EcoR1 cut.
I think we could really use this to save people's lives.
I think we can get -- use this to get medicine.
I mean we're putting an insulin gene into bacteria which is making insulin, so you get insulin that way.
you can see that they're really evenly dispersed and they're also there's not a lot of error
that you can see in the results.
So what have you learned from doing this Restriction Enzyme Analysis experiment?
Everything I know about DNA I've learned from this experiment; I didn't know anything about DNA prior to this.
I didn't know about enzymes, I didn't know that we could cut DNA; I didn't know about any of this.
Yes I would love to pursue a career in college involving sciences, especially genetics. Genetics are probably my favorite course.
How can students in Middle and/or High School get involved in research?
Nothing replaces getting into a lab.
And so I think that really requires 'beating the bushes' and going around to various places like Sloan-Kettering
and Cold Spring Harbor Laboratory to try and find people who are willing to take on and mentor young students in what they do.
There are also various programs. I know, for example, at Cold Spring Harbor there is one for High School students;
Partners for the Future and then places like the DNA Learning Center where one can really gain
hands-on experience with this sort of technology
It was a lot of fun; I want to be a biologist now.