The residual gel was extracted again with extraction solution (5% trifluoroacetic acid (TFA), 67% ACN) at 37°C for 30 min and then centrifuged for 5 min.
After centrifugation at 20,000 g for 6 min, the supernatant was collected and the tissues extracted again as before, and the two extractions for each sample were combined and lyophilized.
Then, after 2 extractions with chloroform, RNA was precipitated with LiCl2, extracted again with chloroform and finally precipitated with ethanol.
Then, after two extractions with chloroform, the RNA was precipitated with LiCl2, extracted again with chloroform and finally precipitated with ethanol.
Instead of direct injection or analysis, the surfactant rich phase containing the complexes was treated by nitric acid, and the detected ions were back extracted again into aqueous phase at the second cloud point extraction stage, and finally determined by ICP-OES.
The aqueous fraction after extraction was acidified with concentrated HCl to pH 2 and extracted again with three equal volumes of ethyl acetate.
For the neutral-acid extraction method, the reaction liquid was directly extracted with an organic extractant, then the pH was adjusted to 2, and the reaction liquid was extracted again.
"You can extract it without damaging the tree, and every nine years, you can extract again.
"Because of the number of incidents we have been forced to report, sadly, in recent weeks, we are less likely to show that kind of extract again," said Chris Cramer, managing director of CNN International.
A "new $10 bill" remembered 50 years after it was withdrawn from his father's wallet, in Ancestors, is extracted again, in a repeat of the same scenario, 20 years later, in "The Front and Back Parts of the House", from Billie Dyer.
