The clones of E. coli transformed as above described were transferred from selective solid medium to 200 µL of liquid medium LB in 96-well plates with the robot colony picker QPIX 450 (Molecular Devices, LLC, California, USA).
Samples were taken at an interval of 24 h. 4 L preliminary seed broth (prepared under the same conditions as previously described) was transferred into a 50-L bioreactor containing 40 L PDB; secondary seed was cultivated at 28 °C for 24 h (agitation speed of 150 rpm and aeration rate of 1.0 VVM).
The layers of the epithelium, obtained as previously described, were transferred into a tube where they were dispersed mechanically by forcing them through a pipette.
Trypsin-digested bone matrix. Fifty samples of rat bone matrix, demineralized as described, were transferred to a solution of trypsin (lyophilized trypsin, 180 U/mg), containing 2 mg of enzyme per ml of 0.1 M phosphate buffer, pH 7.6.
E nzyme digestion. α- Amylase-digested bone matrix. Fifty samples of bone matrix, prepared as described, were transferred to a solution of crystalline amylase (α-amylase 840 U/mg). 1 The solid to solution ratio was 0.1 gm of matrix to 10 ml of a solution of 35 mg amylase per liter of 0.2 M NaH2PO4 buffer, pH 5.4.
At the end of the infection period, 500 μl complete RPMI medium was added, and cells were centrifuged for 1 min at 3,000 rpm and reaggregated as described above, before being transferred on a floating filter for 6 days.
All parameters value described above were transferred into digital format using the Arc GIS software.
Protozoan enrichments described above were transferred sequentially 3 times at 7-day intervals to 250-mL portions of CGWM and incubated at 25°C on a gyratory shaker operated at 50 rpm.
Developmentally synchronized worms prepared as described above were transferred onto the drug plates at L4 stage.
1D and 2D gels run as described above were transferred to nitrocellulose membranes.
