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文献和实验Clonality - X Chromosome Inactivation Assay
(1:1:1, v:v:v) (Gibco) 3 M Na Acetate, pH 5.2 (Life Technologies) Isopropanol Glycogen, 10 mg/ml (GenHunter) 70% ethanol Buffer 1 (50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 50 mM NaCl) Hha I restriction enzyme (Life Technologies) REact 2 Buffer
He Tube Mating with MSCV recipient
. 3. Set up Dilutions: Set up dilutions as shown below, in 5 ml LB+0.2% rhamnose. Use 50 ml Falcons for this as they have good aeration and you get much better growth. Place these dilutions at 30℃ shaking for 2 hours. pMSCV: 25 μl 5ml 50 μl 5ml
Sephadex G-50 spun column purification
. 1) Plug a 1ml syringe barrel with siliconized glass wool and fill to the top with a sterile suspension of Sephadex G50 equilibrated with H2 O. 2) Once filled place the syringe barrel into a 10ml centrifuge tube and spin at 3,000g
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