Chloroplast DNA isolation/mutant detection?

[Stephanie Nelson]

Hello!

One of my former professors is looking for a protocol to use in teaching
a laboratory in plant physiology.  The lab manual that he is using has
an experiment in which the students isolate chloroplast DNA and search
for mutants using RFLP analysis.  One problem:  the protocol calls for
an ultracentrifuge, which the college does not possess.

What I would REALLY like is a protocol for isolating chloroplast
DNA which does not require an ultracentrifuge.  If not, then perhaps
could you point me to a simple lab demonstrating mutant detection
in plants?  The college does have a regular centrifuge, gel apparatus,
access to enzymes, etc., but I don't think they have a thermocycler.

Your help is much appreciated.


Stephanie Nelson
Graduate Student
Case Western Reserve University, Dept. of Genetics