anybody isolating tomato DNA outthere????? - (Jan/27/2001 )
I'm having a big trouble in obtaining high-quality genomic DNA from tomato for PCR purposes. I'm following a new protocol which uses a small blender instead of the liquid nitrogen-mortar-pestle system, but the DNA I obtain seems to be somehow degraded. I've also followed another protocol which I used some months ago with potato DNA (with CTAB) that gave great results, but again, tomato is really a big pain on the neck..., the DNA I obtain really looks awful after Eth Br staining in agarose. Can anyone help me with this?, shall I convince everyone in my lab to start using liquid nitrogen again instead of the blender?, how much can the DNA be mechanically damaged by a blender?, any suggestions/new protocols anywhere?
Thanks a lot
You might also want to try an alternative to Ethiudm Bromide, since it cuts up DNA, esp. long strands. I've tried dilute crytal violet with 1X TAE and/or 1XTBE and it works well. You can actually see the DNA as is runs through the agarose.