For RNA extraction, samples were extracted three times with 20 ml of 65°C phenol (Sigma P4682), 20 ml of Phenol∶Chloroform∶Isoamyl Alcohol (25∶24∶1) (Sigma P3803), and finally, 20 ml of Chloroform∶Isoamyl Alcohol (Sigma C0549), after which the final aqueous layer was removed to a fresh centrifuge tube and precipitated with 2.5 volumes of 100% ethanol (Sigma E7023).
The decline in cell survival in the "CDM" and "CDM2" (water extraction) samples with increasing extract concentration was effectively identical, with zero survival at the highest concentration.
At the time of lipid extraction, samples of bacterial pellets were removed and extracted using the same phospholipid extraction procedure.
For mRNA extraction, samples were frozen in liquid nitrogen immediately after surgical removal.
After extraction, samples (99 μL) were injected into a XBRIDGE C18 column (Waters, Spain) and were scanned by an UV detector at 220 nm with gradient elution.
A simple yet efficient strategy is urgently desired to prepare smart metal-organic framework nanocomposite for highly selective capture and separation of natural flavone from complex extraction samples.
After standing in ice for 10 min, the extraction samples were centrifuged at 5,100 rpm and 4°C for 15 min, and the aqueous layer and precipitates were recovered.
In lysimeter and soil extraction samples, nitrate and ammonium were found at the highest concentrations, while results from microdialysis revealed that the pool of plant-available amino acids was contributing most to the total N pool tested.
For RNA extraction, samples were immediately processed after collection.
Before extraction, samples were surface-cleaned with 10% bleach, 70% ethanol and 254 nm UV radiation.
After 2 min for extraction, samples were introduced into the flow cytometer and fluorescence measured as previously described [22].
