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Seeking method for fatty acid content determination in plant seedsand seedlings

Seeking method for fatty acid content determination in plant seedsand seedlings - Protocols and Methods Forum

Seeking method for fatty acid content determination in plant seedsand seedlings - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 08-03-2009, 03:17 AM
Paul smith
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Default Seeking method for fatty acid content determination in plant seedsand seedlings



Hello,

Not being a chemist, I am seeking a detailed protocol for
extraction and GC analysis of "free" fatty acids and fatty acids in
triglycerides (TGAs)in Arabidopsis seeds and seedlings (time-course
experiments). This is tolook quantitatively and qualitatively at the
amount and composition of TGAs in seeds of a mutant vs. wildtype and
their changes during germination and seedling establishment.

None
of the methods I have seen in the literature is detailed enough for
someone inexperienced to follow without assistance of an expert (none
available where I work). I am also concerned about the safety aspects
with all the dangerous organic chemicals involved. I would be very
grateful to any expert that may provide me with a detailed protocol
that can safely be followed.

I would also appreciate that
details are included for special glassware &equipment that is
required for the procedures. Please note that I do have access to a
Agilent 6890N GC machine with an automatic injector and a 30 m HP-5 30
m column, suitable for FAs.

Thank you!

Paul Smith

[Only registered users see links. ]



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  #2  
Old 08-04-2009, 08:24 AM
StewJW
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Default Seeking method for fatty acid content determination in plantseeds and seedlings

Some commercial lit. can contain some useful info particularly the
right type of GC column to use, although I would search the
sicientific lit. eg

[Only registered users see links. ]

SGE recommend their BP21- Polyethylene Glycol (TPA treated) or a
BPX70-70% Cyanopropyl polysilphenylene-siloxane for the separation of
fatty acid methyl esters. There is also a cross-reference guide to
other manufacturer columns. Fisher UK who I work for also supply
Thermo Hypersil's Trace TR-WAX GC column for FA separation. Make sure
you get the right column for the job is the message, its a competative
field so you should be able to haggle a good price.
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