question about LEAFY

[Gijs Grob]

Hello, I'm a PhD-student working on the phylogeny of Amorphophallus
(Araceae) and in search of a nuclear gene for my phylogeny I've tried the
MADS-box gene LEAFY.
>From Frohlich and Meyerowitz (1997) Int. J. Plant Sci. 158(6 suppl):
S131-S142 I obtained the following universal primers

LFsxl-3
5'-GCC GGC IMG IGG IAA RAA YGG IYT IGA-3'

LFtxr
5'-CCT GCC IAC RTA RTG ICK CAT YTT TIG GYT T-3'

which should amplify a part containing an intron of the presumaby single
copy gene.
However in our Araceae these primers failed either to produce any product,
or to produce a single product or to produce a single product, which
survived boosting. We've tried various buffers and  annealing temperatures,
but so far without success.
What I would like to know is whether there are other people using this gene
and whether you've succeeded in amplifying and sequencing this gene. Or
maybe one of you has designed primers that do work in many plants and which
are better than the two primers mentioned above. Or if all your experiences
with LEAFY are negative, whether you have suggestions for another nuclear
gene I could use on the intrageneric level.

I hope some of you can help me.

Many thanks in advance,

Gijs Grob



___________________________
Gijs Grob
Ph.D-student
Nationaal Herbarium Nederland
Leiden University
P. O. Box 9514, 2300 RA, Leiden
The Netherlands
Tel: (+31-71) 5273583
Fax: (+31-71) 5273511
Grob at nhn.leidenuniv.nl