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Example sentences for: mgcl

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Nuclei were prepared from HeLa cells by standard procedures [ 35] and resuspended in 10 packed nuclei volumes of buffer A (10 mM Hepes (pH 7.9) 1.5 mM MgCl 2 , 10 mM KCl, 0.5 mM DTT, and 0.5 mM PMSF).
After incubation on ice for 12 min, MgCl 2 was added to a final concentration of 3 mM and incubation continued for an additional 5 min.
First strand cDNA synthesis was performed at 42°C for 30 min in a 20 μl reaction containing: 5 μg RNA, 500 nM NEO A primer (5'-ATTGTCTGTTGTGCCCAGTCATA), 20 mM Tris-HCl (pH 8.4), 50 mM KCl 2.5 mM MgCl 2 , 10 mM DTT, 400 μM each dNTP, and 8 units Super Script II reverse transcriptase (Gibco-BRL).
PBB was freshly prepared in 24 ml aliquots composed of 20 ml A-150 (20 mM Hepes, 150 mM KCl, 10 mM MgCl2 and 1% glycerol), and 2 ml each of phosphate buffered saline supplemented, respectively, with 1% Triton X-100 and 1% NP-40.
For each set of heterodimers, 260 μg of p51 was mixed with 660 μg of p66 in the buffer containing 50 mM Tris HCl, pH 7.8, 60 mM KCl and 5 mM MgCl 2 . The rationale for using a 1:3 ratio of p51 to p66 was to saturate the His-tagged p51 with the non-tagged p66, ensuring heterodimer formation and eliminating excess p66 during IMAC purification.