Do you really want to be a part of a trend toward a consumer environment where one master corporation has the potential to set prices and even give its own products preferential treatment in a system used by basically everyone?
KLM was eventually awarded the privatisation of the company, which restructured its debts and made a master corporation agreement with the Dutch airline that bought 26% of the shares, becoming the largest single shareholder since then.
All markers were amplified in 25-μL PCR reactions including: 0.5 μL forward and reverse primers (5 μM), 0.5 μL spermidine (4 mM), 2 μL total DNA, and 5 μL GoTaq Green Master Mix (Promega Corporation, Madison, Wisconsin, USA).
PCR was performed in a final volume of 10 μL, containing 5 μL of 2× GoTaq Hot Start Colorless Master Mix (Promega Corporation, Madison, Wisconsin, USA), 1 μM of each primer, and 60 ng of template DNA.
Real-time quantitative PCR was used to assess MPC expression of selected genes, as listed in Table 1, using the RTReal-Time™ SYBR Green/Rox PCR master mix (SABiosciences Corporation, Frederick, MD, USA) according to the manufacturer's instructions.
Amplification reactions for SP_0048 were performed in total volumes of 10 µL containing 10 ng of template DNA, 0.2 µM of each primer, 0.2 mM of each dNTP, and 5 µL of GoTaq Master Mix (Promega Corporation, Madison, WI).
PCR amplification for the trnL-trnF intergenic spacer was performed in 25-μL reactions using 0.1 μM of each forward and reverse primer, 1 39 ng of genomic DNA, 11 μL of nuclease-free water (Life Technologies), and 12.5 μL of EconoTaq PLUS 2× Master Mix (Lucigen Corporation, Middleton, Wisconsin, USA).
PCR assays had 1 μL of template (5 μL for cerebrospinal fluid) in 24 μL (20 μL for cerebrospinal fluid) of reaction mixture containing 0.2 μM of each primers (Integrated DNA Technologies, Inc., Coralville, USA) and PCR Master Mix (Promega Corporation, Madison, Wisconsin, USA) according to manufacturer's instructions.
First, 10-μL reactions contained 5 μL of Master Mix (Promega Corporation, Madison, Wisconsin, USA), 0.5 μL of 10 mg/mL bovine serum albumin (BSA), 0.25 μL of 10 μM forward primer with the M13 tail, 0.25 μL of 10 μM reverse primer, 3 μL of H2O, and 1 μL of template DNA.
PCR was conducted in a final volume of 25 μl consisting of 1 μl cDNA, 1 μl of 10 pM of each primer (forward and reverse), and 12.5 μl PCR master mix (Promega Corporation, Madison, WI, USA), the volume was brought up to 25 μl using sterilized, deionized water.
The 8-μL PCR reactions contained 4 ng DNA, 6.25 nM HEX-labeled or 9.40 nM FAM-labeled M13 primer (TGTAAAACGACGGCCAGT), 0.075 μM M13-tagged forward primer, 0.3 μM reverse primer, 4 mM MgCl2, 4 μL GoTaq Colorless Master Mix (Promega Corporation, Madison, Wisconsin, USA) including 200 μM of each dNTP and 1 unit Taq DNA polymerase, and H2O.
