Phosphate and sulfate assay extraction from plant
Instrument: Precellys® 24
Sample: Snap-frozen plant tissue, leaves, roots, etc., 30-50 mg
Protocol:
- Weight 20-70 mg of frozen tissue in a 2 ml hard tissue homogenizing CK28 (Cat. No 03961-1-002.2) tube and place on ice
- Add two 2.8 mm ceramic beads and 500 µl 1% acetic acid
- Process in Precellys 24 two times.
- Speed: 5000 rpm Cycle: 3 x 45 sec (break between runs 5 sec)
- Break: 15 min on ice for assays.
- Spin at 14 000 rpm for 15 min at 4ºC, transfer supernatant to fresh tube, spin 15 min at high speed again, transfer one more time.
- Once the extracts are absolutely clear, they are ready for assays.
Results:
Biological space is entirely separated from the mechanical parts preventing any inside contamination. Up to 24 samples can be homogenized at the same time and in less than five minutes. When the protocol is set up and validated, the sample preparation process remains the same with no bias in analysis, on a time basis, or between operators. Moreover, individual sealed tubes prevents cross contamination between sample preparations.
from the Molecular Physiology Laboratory
