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Purified RNA and RT-PCR reagent mix were combined and mixed utilizing the action of the diaphragm valves.
For each real time PCR reaction, 1 ul cDNA, 1 ul gene expression assay, and 10 ul TaqMan gene expression master mix were combined with water in a well on the reaction plate for a total volume of 20 ul.
Briefly, TaqMan® Universal PCR Master Mix Applied Biosystemss, Foster City, CA) and TaqMan® SNP Genotyping Assay Mix were combined with 5-10 ng of genomic DNA.
An IFC controller HX was used to prime the fluidic array with control line fluid, and then the samples and assay mix were combined in the inlets.
RNase-free water, Onestep RT-PCR buffer (1×), 400 μM dNTPs and Onestep RT-PCR enzyme mix were combined with gene specific primers (0.6 μM) to amplify 2.0 μL of template RNA in 50 μL reactions.
The test assay, the reference assay, the DNA sample and TaqMan® Master Mix were combined according to manufacturer's instruction and run on an Applied Biosystems 7500 Real-Time System.
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White rum, coconut liqueur, blue Curaçao, grenadine syrup and a dash of sour mix are combined to create this magic, which is potent and not too sweet, tasting lightly of coconut touched with citrus and winey pomegranate.
In some cases minor groups (A3, B1, B3, C1, C2, C3, Mix) are combined and referred to as "Others".
The RNA-primer mix was combined with M-MLV buffer (50 mM Tris-HCl pH 8.3, 75 mM KCl, 3 mM MgCl2), 10 mM DTT, 0.5 mM dNTP and 0.5 mM M-MLV reverse transcriptase (Invitrogen Inc., Carlsbad, CA).
In preparative PCR, 646.75 μL of master mix was combined with 3.25 μL Taq polymerase.
Seventy microliters of this completed master mix was combined with 5.5 μL of collected DNA in eight separate vials.
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