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PCR mixtures were prepared in a 50 μl volume containing PCR Gold buffer 1× (Roche, Barcelona, Spain), 2.5 mM MgCl2 (Roche), 0.8 mM dNTPs mix supplying 0.2 mM of each deoxinucleotide triphosphate (Applied-Biosystems, Barcelona, Spain), 0.3 mM of forward and reverse primers (Life Technologies, Madrid), 2 U of DNA polymerase (Biotools, Madrid), and 20 200 ng/μl of genomic DNA template.
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From −21 d to calving, cows received the same close-up diet (1.52 Mcal of NEL/kg of DM) and were assigned randomly to treatments (CON, SMA, REA, or MIX) supplied as top dresses.
qPCR was carried out using the Bio-Rad iQ SYBR Green super mix supplied by Roche Molecular Biochemicals.
Second-strand cDNA was synthesized with the DNA polymerase mix supplied by the kit to provide double-stranded DNA template for in vitro transcription.
Droplets were injected into the sorting chip at a flow rate of 30 μL/h and were separated by the oil surfactant mix supplied at a flow rate of 500 μL/h.
For the past five months the centres have been feeding children with nutritious meals – supplemented with protein mix – and supplying play materials.
In other words, given exogenous demand, MESSAGE selects the energy mix that supplies it at least cost [19, 20].
In treatment G CMvw, vermicompost and commercial media mix was supplied with vermiwash and in G CMinor, chemical fertilizer was applied and germination trays were filled with 20 ml of mixed substrate as per treatments in Table 4.
During the electroless silicon etching process, we did not use any artificial stirrer or sonicator to mix and supply the solution near the etching surface or to destroy the initial air bubbles inside the microcavities.
Fast Sybr Green master mix was supplied by AB Applied Biosystemm, USA).
PCR master mix was supplied by New England Biolabs (Schwalbach, Germany).
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