Reverse transcripts were used as templates for analysing gene expression levels using AmpliTaq Gold PCR Master mix (Applied Biosystems) and a PX2 thermal cycler (Hybaid) according to the manufacturer's instructions.
The reaction was incubated at 25°C for 5 min, followed by 42°C for 30 min, and ending with 85°C for 5 min. After reverse transcription, the cDNA reaction was diluted to a volume of 50 µl with nuclease-free water and used as a template for qPCR analyses.
They conclude that their findings provide the basis for prioritizing pollution-control strategies in the Yangtze River Delta and provide a template for comparable analyses elsewhere.
This approach allows the implementation of non-proofreading, highly processive polymerases that limit costs yet generate substantial quantities of template for downstream analyses.
After centrifugation of the C. difficile DNA containing solutions, 125 μL of supernatant was collected and stored at –20°C as a template for PCR analyses.
The purified DNA was used as template for qPCR analyses with a Bio-Rad iCycler and Sybr Fast Q-PCR mix (Kapa Biosystems, Wilmington, Massachusetts, USA).
Our findings, in combination with plausible emissions reduction estimates and their costs, provide the basis for prioritizing pollution control strategies in the YRD and provide a template for comparable analyses elsewhere.
Due to consistent results from unconstraint tree reconstruction and hypotheses testing, we took the topology reflecting the ' glob2/3 duplication-in Sophophora subgenus' scenario as template for subsequent analyses of globin sequence evolution.
As template for these analyses, we used control and PTU heart cDNA and we targeted the cardiac MHC RNA expression, a condition associated with significant change in MHC gene expression [ 31].
PL: designed and performed the BLAST analysis in Table 2. SRZ: designed and executed the mixed-model analysis used in Table 6 and as the template for Community Collaboration analyses.
