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文献和实验Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library
B. Culture/expand cells. We have obtained excellent results with 2 x 107 cells per library transfection. C. Trypsinize cells that are at >80% confluency, and transfer 2 x 107 cells to a 15 ml sterile screw topped tubes.
ChIP protocol for X. laevis Lens1/FoxE3 enhancer
(1) Remove vitelline membrane from stage 21-23 embryos (n = 300). Collect head tissues (about anterior 1/4 or 1/3 of the embryo) in 1x MBS. (2) Fix the tissues in 0.5x MBS/1% formaldehyde (4-5 ml in a screw cap glass vial) for 15 min
by centrifuging the gradient at 100,000 x g (28,000 rpm)for 3 h at 4℃ in a SW-28 rotor. 5.In the cold room,with a Pasteur pipette carefully harvest the off-white band of Golgi membrane at the 0.25 M/1.1M sucrose interface and the off-white band of ER (ER)membrane
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