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Aliquots of SimReg1 fusion protein in storage buffer were stored at - 80°C, or used immediately in DNA-binding assays.
Samples in SM buffer were stored at 4°C until further use.
MACS-sorted prostate cell lysates in RLT buffer were stored at -80C for no more than a month.
Final protein preparations in Laemmli buffer were stored at -20°C until use. 3 μg of DNase-digested total RNA was used for oligo (dT) primed first strand cDNA synthesis with SuperScript amplification (Invitrogen) following manufacturer's suggestions.
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As 'input', 40 μl lysate +25 μl 4× loading buffer was stored at −20°C.
After removal of cell debris by centrifugation at 250 ×g for 10 min, the supernatant was centrifugated at 20,000 ×g and 4°C for 20 min and the pellet, resuspended in SPG buffer, was stored at −70°C.
Buffers were stored at 4°C until use.
All stock buffers were stored at 4°C and were kept for 1 month.
Treated cells were lysed in RIPA buffer and lysates were stored at −80 °C.
The pellet was homogenized in 4 volumes of glycerol phosphate KCl buffer, and aliquots were stored at −80 °C.
Elution was performed in 20 μL of Elution Buffer and samples were stored at −20°C until sequencing started.
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