Despite these differences, we demonstrate that minute quantities of synthetic Tau fibrils can be amplified in vitro and in crude brain extracts of AD tissue.
A single copy gene was amplified in parallel to normalize the quantity of the input DNA.
Briefly, cDNA was synthesized from 2 μg of total RNA using the TaKaRa RNA PCR Kit (TaKaRa), and the mRNA for UBQ10 was amplified in parallel reactions to normalize the cDNA quantity added to each reaction [ 62].
A challenge arises in that it is not the total residual confounding in Model 1 (the quantity being sought) that is amplified in Model 2, only the fraction of that residual confounding that remains after introduction of the introduced variable.
To normalize the quantity of total mRNA, GAPDH mRNA was amplified in parallel in different tubes.
cDNA for UCK2 and ACTB were amplified in a multiplex reaction for the purpose of increasing the quantity of the target cDNA prior to quantitative real-time PCR.
To normalize the quantity of the input DNA, a single copy gene was amplified in parallel as well.
Five different quantities of the target sequence (10,000, 100,000, 150,000, 200,000, and 1,000,000 molecules) were amplified in the presence of dilution series of the competitor sequence.
