Cells were scraped, collected, and rocked on ice for 30 min.
The samples were scraped, collected and centrifuged at 10,000 g for 30 minutes.
The cells were scraped, collected and lysed for 30 min.
Cells were washed twice with ice-cold PBS, scraped, collected in a microcentrifuge tube, and then centrifuged.
Cells were scraped, collected, triturated with a 257/8 gauge needle, and the homogenate was centrifuged for 10 min at 10,000× g.
Cells were then scraped, collected and transferred at a split ratio of 1 : 3 to a new plate on Mito-MEFs in hESC medium.
The cells were washed, scraped, collected, and centrifuged at 45000 × g at 4°C for 1 h to yield the whole cell extract, as previously described [ 9].
After washing, cells were scraped, collected at 150 g for 5 min, resuspended in PBS (200 µl) and solubilized by addition of 20 volumes of chloroform/methanol (2 1 v/v).
Cells were washed with PBS, scraped, collected by low-speed centrifugation, and lysed with lysis buffer (6.25 mM Tris [pH 6.8], 1% SDS, 10% glycerol, 6 M urea, 0.01% bromophenol blue, 0.01% phenol red) for 30 min on ice.
Cervical scrapes collected from the same cervical cancer patients (n=59) also revealed similar results suggesting that the telomerase activity of exfoliated cells reflects that of the lesions or tumour tissues.
