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The frozen tissue blocks were then placed on dry ice and subsequently stored at −80 °C until further use.
The tissue blocks were then placed on dry ice and subsequently stored in the -80°C freezer until being sectioned for histological analysis.
Needle core-biopsies (0.6 mm) from the corresponding areas on the paraffin-embedded tumour blocks were then placed at pre-specified coordinates in recipient paraffin array blocks using a manual tissue arrayer (Beecher Instruments, Sun Prarie, WI, USA).
Both blocks were then placed into a petri dish under cotton pellets soaked with room temperature distilled water and placed in an incubator at 37°C for 24 hours at 95% humidity.
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The samples were then placed onto blocks of dry ice until the tissues and OCT were frozen to a solid white.
Brains were then placed in plastic blocks in insulin OCT (TissueTek) and frozen.
Tissues were then placed into tissue block molds filled with paraffin.
They were then placed in a blocking solution [3% normal goat serum (NGS) (Vector Laboratories, Peterborough, UK) in PBS with 0.3% triton-X (PBS-T)] for 1 h to reduce background staining.
Samples were then placed on the magnetic block for 2 min, and the supernatant withdrawn.
100ul sheared chromatin aliquots were then placed on 95°C heat block for 10 minutes.
The tissue block was cut into 10 μm thick sections, which were then placed on glass slides and allowed to dry overnight at room temperature.
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