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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
Single tube confirmation PCR protocol
Polymerase. - Add 35 µl of the PCR mix to each of the 20 PCR tubes that already contain the 15 µl of primers and template. 110 µl 5 µl 10 x Taq buffer(see below) 11 µl 0.5 µl 20 mM dNTP's (0.2 mM) 11 µl 0.5 µ
He Tube Mating with MSCV recipient
the dilution with the OD closest to 0.1 (this is true for recipient AND donor cultures. You want an actively growing culture for optimal mating) 5. To set up tube mating, you want a 1:1 ratio of recipient:donor. Determine which dilution of the MSCV dilutions
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