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Co-Expression of human DNA primase p49-p58 subunits

) •3mM BME (to prevent disulfide formation,Ni beads are compatible w/low concs.of BME,but NOT DTT.) Wash buffer: •1% Triton X-100 •50mM Tris,pH 8.0 •300mM NaCl (Qiagen recommends at least 300mM NaCl to prevent ionic interactions w/Ni-NTA beads.NaCl

Purification Techniques of Subcellular Compartments for Analytical and Preparative Purposes

of subsequent steps. Breaking vegetative amoebae by shear stress with a steel ball cell cracker preserves the integrity of subcellular organelles and in particular that of lysosomes, the rupture of which is very deleterious to further purifications.

RNA-biotin based pulldown assays for the detection of siRNA targeted genomic regions and siRNA directed histone modifications (PROT32)

and incubate on the rotating platform at 4°C overnight;Add 20µl Protein A/Salmon sperm sephrose beads to each sample and incubate at room temperature for 15 minutes on a rotating platform (comment 3);Centrifuge the samples containing the Protein A/Salmon spern