The phrase "afterwards by adding" is correct and usable in written English.
It can be used when describing a sequence of actions where something is added after an initial step or event.
Example: "The recipe requires you to mix the ingredients first, and afterwards by adding the spices, you enhance the flavor."
Alternatives: "subsequently by including" or "later by incorporating".
The is gradually increased to afterwards, for example, by adding to every 1 second.
The mixture was vortex mixed for 10 s followed by incubation at 50°C for 15 min. Afterwards, the reaction was terminated by adding 2 mL of ice-cold distilled water and making up to 5 mL with acetonitrile.
Afterwards, silylation was carried out by adding 100 µl of N-methyl-N- trimethylsilyl -trifluoroacetamide (MSTFA) with 1% trimethylchlorosilaN-methyl-N- trimethylsilyl -trifluoroacetamideample aN-methyl-N- trimethylsilyl -trifluoroacetamide
Afterwards, the reaction was quenched by adding its volume of reducing electrophoresis sample buffer.
Afterwards ubiquitination mixture was quenched by adding 12.5 μL 2 × non-reducing gel loading buffer and heated for 5 min to 80 °C.
Afterwards, the reaction was stopped by adding 50 μL 1M Na2CO3, the total reaction time noted and the absorption measured at 405 nm (reference wavelength 620 nm).
Afterwards, cell lysis was performed by adding 10 μl of lysis-solution (Evotec) and 90 μl lysis buffer (Evotec) to each well and incubating the suspension for one hour at room temperature in the dark on a shaker.
Afterwards, polar samples were derivatized by adding 120 μL of 15 mg mL-1 of methoxyamine HCl in pyridine, sonicated until all crystals disappeared and incubated at 50°C for 30 minutes.
Afterwards, the slides were neutralized by adding Tris buffer (pH 7.5), stained with 30 ml of ethidium bromide (Sigma-Aldrich, St Louis, MO, USA) (20 mg/L), and then covered and stored in sealed boxes at 4°C for further analysis.
