siRNA Electroporation Buf. 1.5

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

the 3 hour incubation Dynabeads™ M-280 Streptaviden magnetic beads (7x107 beads, ~100µl/sample) are added to the respective samples (note 1);5.The resultant siRNA/Flag complexes are then eluted with magnetic bead binding, the solution can be discarded

Transformation of E. coli by Electroporation

absolute ethanol. Ice 15 min. 3. Centrifuge at >12,000 x g for 15 min at 4° C. Carefully decant the supernatant. 4. Wash the pellet with 1 ml of 70% ethanol. Centrifuge at >12,000 x g for 15min at room temperature. Remove the supernate. 5. Air

Electroporation of ES cells

for each 10 cm plate). 7. Determine the cell density (haemocytometer) and dilute with PBS to the required density for electroporation. We regularly electroporate at a relatively high cell density: 7x106 cells/ml (this number varies between different labs