4 replies to this topic

[Mad researcher]

I want to extract DNA from fin clips (fish).
I would follow the GEN ELUTE MAMMALIAN DNA MINIPREP KIT.

How do you suggest i mince the fin clip and analyze for DNA thereafter?

It would be nice if someone can suggest a fast method to do it. If you suggest liquid nitrogen then please tell me how to proceed with that.

[prabhubct]

Mad researcher, on 02 October 2012 - 05:49 AM, said:

I want to extract DNA from fin clips (fish).
I would follow the GEN ELUTE MAMMALIAN DNA MINIPREP KIT.

How do you suggest i mince the fin clip and analyze for DNA thereafter?

Mincing and then extracting DNA by kit will ok. But you have to look for specification for fish (it's not mammal u know it).

[bob1]

The usual way of doing these sorts of things (usually used for genotyping) is to digest in proteinase K for a couple of hours then proceed with a crude DNA prep using salt and ethanol.  I don't know how compatible this will be with the kit you want to use.
Grinding in nitrogen is pretty fast - you need to have some mortar and pestles or microuge tube grinders.  Pour a little LN2 into the vessel, enough that the sample will freeze, then attack it with some vigour.  Once ground, either scrape into an appropriate tube or add the lysis buffer and transfer to a tube.  If you have any liquid (such as water or other buffering solution) around the tissue, it will form a solid lump which will be very hard to grind up, so make sure you dry off the fins before grinding.

[Mad researcher]

can't i just mince the tissue using a sonicator in a eppendorf tube?

[bob1]

A sonicator won't work particularly well on whole chunks of tissue.  You would be better off using a polytron homogenizer or perhaps a dounce.