Lipid Profiling Extraction Method for Arabidopsis 1xbet online games login

A .pdf of 1xbet online games login method, including shipping directions, can be found here.

NOTES: Use 1xbet online games login tubes with Teflon-lined screw caps. Use a Dounce-type homogenizer – ground 1xbet online games login to ground 1xbet online games login . Use 1xbet online games login Pasteur pipets for transferring seeds/solvent.

Volumes 1xbet online games login doubled for larger samples or ease of homogenizing.

1) Weigh 1xbet online games login , using a microgram balance. Lyophilizing first may be best for consistent dry weight.

2) Drop seeds into 1.0 1xbet online games login 75ºC isopropanol with 0.01% BHT. Heat 15 minutes. Let cool. (Hot isopropanol inactivates phospholipase enzymes.)

3) Transfer to homogenizer; homogenize thoroughly. Transfer to 17 1xbet online games login glass tube for extraction. Rinse homogenizer with 1.0 1xbet online games login chloroform and 1.0 1xbet online games login methanol to recover all the seed parts/lipids; add to isopropanol/seed mix in tube. Add 0.8 1xbet online games login water to solvent/seed mix. Shake well – should be one phase.

4) Add 1.0 1xbet online games login chloroform and 1.0 1xbet online games login water. Shake well. Centrifuge to split phases.

5) Remove lower layer (1xbet online games login and lipids). SAVE to clean glass tube.

6) Add 1.0 1xbet online games login chloroform, shake, and centrifuge. SAVE this lower layer in same tube as first.

7) Add 1.0 1xbet online games login chloroform, shake, and centrifuge. SAVE this lower layer in same tube as above.

8) Add small amount (0.5 1xbet online games login ) 1 M KCl to chloroform/lipids. Shake well; centrifuge. Remove thin water layer on TOP and discard.

9) Add small amount (1.0 1xbet online games login ) water to chloroform/lipids. Shake well; centrifuge. Remove thin water layer on TOP and discard. (This backwash removes any proteins and carbohydrates that may have been carried through the extraction.)

10) Dry to small volume. Fill tubes with nitrogen, store at -20°C (freezer). When you are ready to prepare for shipping, evaporate completely under nitrogen gas or a speedvac, and redissolve in about 1.0 1xbet online games login chloroform. Transfer to 2.0 1xbet online games login clear glass vial with Teflon-lined screw cap (for example: clear glass; 2 1xbet online games login ; solid PTFE-lined cap; 1/2 dr., Fisher Scientific catalog #03-391-7A, Thermo Scientific No.: B7800-1). Evaporate the solvent before 1xbet online games login . ALWAYS 1xbet online games login us know before you send your samples: mrroth@ksu.edu