Individual DNA samples were pooled (50 ng per individual) and subsequently concentrated using a DNA concentrator Eppendorf 5301.
The aqueous phase was collected after centrifugation at 14,000 rpm for 10 minutes and was subsequently concentrated into 60 μL of DI water after vacuum centrifugation.
The fractions containing proteins (MBP-ΔN597 or MBP-ΔNCP69) were confirmed by SDS PAGE, and were subsequently concentrated using 10 kDa MWCO Vivaspin Sample Concentrators (Sartorius Stedim Biotech).
PolyA RNA was subsequently concentrated by ammonium acetate ethanol precipitation and concentrations were determined using the Qubit fluorometer (Invitrogen, Carlsbad, CA) and the Quant-iT RNA Assay Kit (Invitrogen, cat#Q33140).
After centrifugation, the solution was subsequently concentrated with centricons (Millipore, MA, USA) up to about 100 μL volume, and then the extraction proceeded according to the handbook of the QIAquick DNA Purification Kit (Qiagen, Hilden, Germany).
Purified DBLs were subsequently concentrated by means of Macro- and Micro-sep concentrators.
The eluted and neutralized samples were combined, diluted to 20ml with MilliQ water and subsequently concentrated to the original starting volume using centrifugal concentration tubes [Vivaspin 20, Sartorius AG, Goettingen, Germany: Code no VS2021].
The protein was cleared of imidazole using dialysis with phosphate buffered saline and subsequently concentrated using a 9 kDa cut-off protein concentrator according to the manufacturer's instructions (Thermo Fisher Scientific, Rockford, IL, USA).
Viruses were subsequently concentrated from conditioned media harvested 48 and 72 h after transfection by ultracentrifugation at 25,000 rpm in a Beckman SW28 swinging bucket rotor.
Mr. Kluge, 87, subsequently concentrated on filmmaking, restaurant and telecommunications ventures.
