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文献和实验with the better the chances of a successful pulldown. A total of 4.0×106 293 HEK cells (~70% confluent) are plated and 24 hours later transfected with 15µg of a RNA Polymerase II expressing flag-tagged construct plasmid. We generally use Lipofectamine 2000™
DETECTION OF ß-GALACTOSIDASE AND ALKALINE PHOSPHATASE ACTIVITIES IN TISSUE
background. 4. Incubate tissue in X-Phos/NBT Detection Buffer for 15' at room temperature. XPhos/NBT Detection Buffer (Buffer 3 as described for Genius kit by Boehringer-Mannheim) 100 mM Tris-HCl, pH 9.5 100 mM NaCl 50 mM MgCl
RNAs;Following the 3 hour incubation Dynabeads™ M-280 Streptaviden magnetic beads (7x107 beads, ~100µl/sample) are added to the respective samples (note 1);The resultant siRNA/Flag complexes are then eluted with magnetic bead binding, the solution
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