The pellet was then gently resuspended in high-salt isolation buffer, and the above medium- and high-speed centrifugation steps were repeated to produce a final enriched mitochondrial pellet.
Therefore, the controlled-release discs enable the user to control the pH without any additional buffer and the osmolarity of the medium was dramatically reduced from 0.78 OsM in the medium with 0.2 M MOPS buffer to 0.44 OsM in the medium without MOPS buffer.
Total cell lysates were prepared with RIPA buffer 48 72 h later, and the medium was centrifuged at 13,000 rpm for 1 min to remove cell debris.
In the beginning of the cultivation with a MOPS buffer concentration of 0.2 M, the pH did not change because of the high buffer capacity of the medium and the relatively low metabolic activity of the microorganisms.
This study related the dissolution enhancement of a TCE DNAPL to the pH buffer capacity of the medium and the type of electron donor used.
Peptone (0.5% w/v) was added to the extract and the medium buffered to pH 7 using potassium phosphate buffer.
Following each taxotere treatment, the cells were washed twice with phosphate buffered saline (PBS, Nissui, Tokyo, Japan) and the medium was replaced with fresh complete medium for an additional 3 days.
Nine factors were selected (the concentration of glycerol, tryptone, yeast extract, PBS buffer, and MOPS in the medium, induction occasion, the concentration of IPTG, inoculation, loading volume), each of which was coded with two levels (Table 1).
Briefly, 2×10 cells were collected and washed with PBS, acidic glycine buffer and RPMI medium.
Pt1 was grown at 20 °CC in 1.25 L photobioreactors in ASPII media as previously described [ 21, 22], and the medium was buffered at approximately pH 8.0 with Tris-base.
