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In this process, a succession of steps with a mutation and local optimization (the method of Broyden-Fletcher-Goldfarb-Shanno [BFGS]) were conducted, and each step followed the Metropolis criterion [24].

Conventional peptide-MHC-tetramer production involves recombinant MHC production, in vitro refolding, biotinylation and tetramerization; each step followed by various biochemical steps such as chromatographic purification, concentration etc.

His6 Cas8′ was purified on an AKTA FPLC with each step followed by SDS/PAGE.

Creep tests were performed by increasing the force stepwise in increments of Fmax/10 each step followed by a 500-s holding period (Fig. 1C).

They were then washed in a series of ethanol (50 – 100%), ten minutes at each step, followed by two additional periods of absolute ethanol (10 minutes each).

The amplification program consisted of five cycles of 1 min each step followed by 45 cycles of 30 s each step.

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The dialogue shows what is so seductive about mathematics, that each step follows inevitably from the previous step, in a way that makes it absolutely beyond doubt or error.

The outline of each step follows.

All QDs were coated with ZnS by adding a total of 0.8 mL Zn OAc 2 at 240 °C in two equal portions, allowing a reaction time of 30 min after each step Following the preparation of alloyed (ZnS x(CuInS2 1−x core particles, shell formation was accomplished by raising the temperature to 240 °C, adding 0.8 mL of Zn OAc 2 in two equal portions and allowing a reaction time of 30 min after each step.

Samples were submitted to 6 freeze-thaw cycles (−20°C/room temperature, with variation from 22 to 26°C), each thawing step followed by NMR analysis.

This is a hill climbing iterative algorithm that requires two steps in each iteration – the expectation (E) step followed by the maximization (M) step.

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