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Working in pairs or individually, students prepare genomic DNA from Canton S or dy frozen flies.
For CHC extraction, groups of 5 frozen flies were shaken in a glass vial with 200 µL of Hexane.
For primary screening, frozen flies were manually transferred to a new flat-bottomed 96-well plate (Daslab, Barcelona, Spain) by inverting the deep-well plate over this new plate.
Heads were separated from the remaining bodies by quickly vortexing frozen flies and collected by filtering them with liquid nitrogen through a sieve with an aperture size of 710 µm (diameter) on top of a sieve of 425 µm (Van Gelder et al., 1995) (https://www.youtube.com/watch?v = Dn2mp9OKhFc).
Heads were then dissected from the frozen flies in each age/experience category.
10 ml frozen flies in liquid nitrogen were gently shaken in a 50 ml-Falcon tube together with 5 ml of glass beads (diameter 4 mm) as described [16].
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Made in Italy, it is flash frozen, flown to Brooklyn and baked in an electric oven on a ceramic hearth.
Although histamine was not detected in any frozen samples, it accumulated rapidly in the previously frozen flying fish meat and once thawed and stored at 25°C.
Once the frozen flying fish samples stored at −20°C for 2 months were thawed and stored at 25°C after 24 h, histamine started to accumulate rapidly (>50 mg/100 g of fish).
Although no histamine was detected in any of the frozen flying fish samples right after thawing, it began to accumulate rapidly after 12 h of storage at 25°C (Fig. 4c).
A volume of 4 ml (2 g) frozen fly heads gave typically 45 mg of total MP with 0.5 mg HsSERT (1 mg DmSERT) purified routinely using an affinity column (Figure 3H).
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