HELP! Tomato genomic library

Kristina Gruden, IJS Kristina.Gruden at ijs.si
Fri Feb 11 03:12:14 EST 1994


Hi!

I have problems with preparing tomato genomic library. We're using Stratagene
system and it works fine with human DNA. So I suppose that the problem is
in the DNA contaminants. I'm using the folowing protocol for DNA isolation:
1. isolation of tomato nuclei (Honda buffer)
2. lysis of the nuclei 
3. some polysacharides are still present, so I use CTAB purification together 
   with phenol-chloroform extraction
4. I precipitate DNA with 100% EtOH and wash it with 70% EtOH and Sodium acetate
pH=5,2

The titer of the library is very low - only 10000 phages/ml.

Any suggestions are welcome

Kristina



More information about the Methods mailing list