FINK%DULRUU51@PUCC.PRINCETON.EDU (10/09/90)
I am working on a project where the main interest is the isolation
and restriction of genomic DNA of cultured human melanocyts.
I would like to look for the genomic situation of the NF1 gene by
restriction and PCR, respectively. The conventional methods of
isolating DNA (ProtK, phenol extraction; high salt) failed because
of the high melanin content of our cultured cells. Even isolated
nuclei (tested under lightmicroscopy) did not allow to isolate pure
DNA. The DNA containing solution is allways yellow or even dark
brown due to the melanin conntent. The DNA is of poor quality for
my investigations because neither restrictionenzymes cut the DNA
efficently nor could-PCR products be obtained. We believe that
the contamination with melanin are responsible for these problems.
Does any body know how to get pure DNA from cultured
primary human melanocytes who produce a lot of melanin ???
mail: Dr. Thomas Fink
Dept. Human Genetics
Albert-Einstein-Allee 11
7900 Ulm/Donau
Germany
phone: 0731 176-3247
BITNET: FINK@DULRUU51