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Following another 48-h incubation, the media was removed and 250 l of RIPA buffer (50 mM Tris, 150 mM NaCl, 0.5% sodium deoxycholate, 1% NP-40, 0.1% SDS, pH 8) was added to each well

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The mixture was incubated at room temperature for 15 min, followed by adding 1 mL of diluted PEG (0.8 mL of PEG + 0.2 mL of TE/Cation Kit components) and incubated at room temperature for another 3 h

However, it is important to note that there are limited studies on either protocols effect on methylation, and no studies on Yaskos utilization of SNP data to drive decision-making about supplements

Gastroenterology 132 , 13441358 (2007)