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文献和实验Clonality - X Chromosome Inactivation Assay
. Pipet upper phase into new tube and discard lower phase. Add 5 µl 3M NaAc. Add 250 µl isopropanol. Add 2 µl glycogen. Vortex briefly. Place on dry ice for 30 min. Centrifuge 14,000 rpm for 30 min. Add 300 µl 70% ethanol. Vortex briefly
.27. Column Buffers Make 500mls equivalent of base buffer: 10ml 1M Hepes pH 7.9 100ml 10% glycerol 5ml 10% NP40 (or 5.0ml 300mM Octyl glucoside) 200ml 0.5 EDTA ------- 115.2ml /5 = 23ml Base 1M KCl2.5M KCl dH20 for 0 KCL 46ml 0 0 154ml = 200
Native Chromatin Preparation and Illumina/Solexa Library Construction
off supernatant completely. 14. Resuspend the cells completely into appropriate volume of T cell isolation buffer. Use 3 mL for 250-300 million cells. 15. Proceed to magnetic separation using an LS column (Step 16).
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