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Scratch wounds were then created using a pipet tip and the wound site photographed digitally at two-hour intervals.
We maintained the droplet above our objective using a pipet tip and slowly translated the substrate below the droplet.
Scratch wounds were generated in the cell monolayer using a pipet tip and each wound was photographed at 0 and 24 hours.
The pellet was then removed using a pipet tip to draw the pellet up the side of the tube and out of the supernatant.
We employ droplets containing as little as tens of picograms of material and simply drag them, using a pipet tip, over a polymer-coated coverslip.
Scratch-assays, where a monolayer of cells is mechanically wounded using a pipet tip, are traditionally used for epithelial cell wounding (Sholley et al., 1977).
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Wounds were made using a p20 pipet tip and cells were given 24 hrs (Hec50 and BT549) or 48 hrs (AN3CA) to migrate into wounds.
After the cells were allowed to attach and reach 80% subconfluency, they were incubated with starvation medium containing 2% FCS for 24 hours prior to further incubation for 2 hours in starvation medium in the absence (control) or presence of Curcumin at concentrations of 10, 20, and 50 μM, before a scratch was performed through the cell monolayer using a yellow pipet tip.
The entire procedure was automated using a pipetting robot.
Following ligation the library was packaged using T7 packaging extracts previously thawed on ice and mixed by stirring with a pipet tip.
Cells were collected using a cell lifter (Thermo Fisher, Waltham, MA, USA), transferred to an Eppendorf tube, passed through a pipet tip five times and incubated on ice for 10 min.
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