DNA Extraction from Fruit tissue

[Hamidrad]

IS there any one who did extraction from pulp or peel of fruits (as kind of banana, apple or orange) ?
I am doing it know but I am confused why sometimes the DNA is extracted and sometimes ... !!

[neja]

Hi

I was working with plant DNA but mostly leafs and roots. Roots are quite challenging.

I used CTAB method which always worked on leafs and most of the time also on roots. When working with roots I noticed that the efficiency of the DNA extraction depends on the season, in autumn it was much less efficient than in spring and summer. I suspect this was because of some substances that are produced later on in the season, like polysaccharides and polyphenols, and it can also be the case in fruits. There are methods developed for DNA extraction from polysaccharides rich material, such as friuit, try to google it. I know for sure that CTAB method is suitable. Always also use phenol purification because this will further remove the polysaccharides.



Another important issue with DNA extraction was a sample preparation. If you are using liquid nitrogen the sample must be really pulverized, no bigger particles must remain. If you grind your samples, which is easier in the case of fruits becasue they are wet, than you also have to grind it untill you don't see particles anymore.


good luck,
neja

[Hamidrad]

Originally Posted by neja

Hi

I was working with plant DNA but mostly leafs and roots. Roots are quite challenging.

I used CTAB method which always worked on leafs and most of the time also on roots. When working with roots I noticed that the efficiency of the DNA extraction depends on the season, in autumn it was much less efficient than in spring and summer. I suspect this was because of some substances that are produced later on in the season, like polysaccharides and polyphenols, and it can also be the case in fruits. There are methods developed for DNA extraction from polysaccharides rich material, such as friuit, try to google it. I know for sure that CTAB method is suitable. Always also use phenol purification because this will further remove the polysaccharides.



Another important issue with DNA extraction was a sample preparation. If you are using liquid nitrogen the sample must be really pulverized, no bigger particles must remain. If you grind your samples, which is easier in the case of fruits becasue they are wet, than you also have to grind it untill you don't see particles anymore.


good luck,
neja

Thanks for your help. So I am trying it. but I should mention that I couldnt find any protocols for pulp or peel of fruits and sometimes I have a good result and another time NO. I dot know whats the problem. I am thinking about If a fruit stored in refrigerator or keep in -20 freezer we lose dna !? ...