8 replies to this topic

[hsbiopeep]

Hey, so I am a Junior in High School and my partner and I are conducting a science experiment for our Biology Advanced class. Our topic is observing the similarities and differences in the DNA of various fruits and vegetables by extracting the DNA and performing gel electrophoresis. We are a bit stuck because we do not know which restriction enzymes would work best for fruits nor vegetables. (We cannot perform PCR, therefore we are using restriction enzymes).

Could someone help us?

[pito]

hsbiopeep, on 31 March 2012 - 09:17 AM, said:

Hey, so I am a Junior in High School and my partner and I are conducting a science experiment for our Biology Advanced class. Our topic is observing the similarities and differences in the DNA of various fruits and vegetables by extracting the DNA and performing gel electrophoresis. We are a bit stuck because we do not know which restriction enzymes would work best for fruits nor vegetables. (We cannot perform PCR, therefore we are using restriction enzymes).

Could someone help us?

You can use whatever you want.

Its true, some* will cut more then others, but it doesnt really matter for what you are doing..
Restriction enzymes are expensive, so I advise you to buy the cheapest ones.. and use those.
(and buy those that you can use: check what buffer you need to use, what temperature, incubation time... I would take these things in concideration ==> also think about "can I mix RE A and B in the same sample/condition + the cost of course)


*hint: use at least 1 rare cutter and 1 that cuts more often/easly.
Check: http://en.wikipedia....triction_enzyme also to know more about the enzymes and about what is important (like co-factors)

[hsbiopeep]

So will it be okay if I use Hae3, Pst1 and Hind3?

[pito]

Sure why not.
Hae III wiill cut a lot, so its possible you will get very small pieces of DNA.

[swanny]

And ask the suppliers if they will give you some enzymes. You'll never know if you never ask...

[hsbiopeep]

pito, on 15 April 2012 - 10:32 AM, said:

Sure why not.
Hae III wiill cut a lot, so its possible you will get very small pieces of DNA.

So is it good to have small pieces of DNA or not?

[pito]

hsbiopeep, on 22 April 2012 - 04:04 PM, said:

pito, on 15 April 2012 - 10:32 AM, said:

Sure why not.
Hae III wiill cut a lot, so its possible you will get very small pieces of DNA.

So is it good to have small pieces of DNA or not?

Well.. it depends...
It all depends on what you want.

What I wanted to say with it is that you will get many small pieces.. so you will have a gel with many pieces.. rather then a few.. It could be more difficult to see differences ...
Also: small pieces means: watch out with the time you let the gel run.. the pieces could migrate off the gel.

[hsbiopeep]

I was given this topic by my biology teacher but I am not sure how I will be able to observe the differences between fruits and vegetables.

Will certain bands only be present in fruits rather than vegetables?

What I am trying to ask is how should I try to observe the differences between fruits and vegetables?

[pito]

hsbiopeep, on 24 April 2012 - 04:14 PM, said:

I was given this topic by my biology teacher but I am not sure how I will be able to observe the differences between fruits and vegetables.

Will certain bands only be present in fruits rather than vegetables?

What I am trying to ask is how should I try to observe the differences between fruits and vegetables?

Its all you can do: use the RE and then watch what it gives on the gel.
I have never done this before, so not sure what to expect.
But indeed: the more the DNA is different, the more difference you will see in the bands...

Just test a few different kinds of fruits and test some vegetables, put them on gel (more then 1 sample per fruit/vege) and check the differences.
Check fruits vs fruits and fruits vs vegetables and vegetables vs vegetables

Also: be sure to test the tomato.. (look up: Nix v. Hedden) and see for yourself whether the court was right or not...
(be sure to try to find a definition on what a fruit or vegetable is..)
(==> also starting from this "definition" you should be able to find out whether there would be a big difference or not and what might be the cause of this difference on DNA level..)