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文献和实验Nucleobond Column BAC DNA Purification for Transgenic Mouse Production
cell pellet in buffer S1: 12 ml (see Note A above) 2. Add the appropriate volume of buffer S2: 12 ml (see Note A above) Mix gently by inverting the tube, and incubate at room temperature or 5 min. Do not vortex to prevent the release of chromosomal
and allow it to settle by gravity. （DO NOT MIX）8. Gently wash the column 3X with 750 l 1X NETS. （Pipette down the side of the column without disrupting the cellulose.）9. Elute the mRNA into a fresh Eppendorf tube by pipetting 650 l of 65 !C 1X ETS directly
, and 20 mMTris-HCl, pH 7.5, sterile filtered. 5. Syringe (30–50 mL) with 0.2-μm pore size filter. 2.3.2. Step 2: IMAC 1. Sample: Pooled Wnt3A containing fractions from Section2.3.1 . 2. HiTrap? Chelating, 1-mL column (GE Healthcare, Piscataway
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