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文献和实验ORNL MICROARRAY HYBRIDIZATION PROTOCOLS
for 10min Immediately neutralize the mix with 5ml of 1M Tris-HCl, pH= 7.5 ・ If only using single label, add 38.5ml of 10mM Tris-HCl, pH=8.0, 1mM EDTA (Volume = 67ml) ・ For dual labels, combine the Cy3 with the Cy5 (Volume = 57ul). Then rinse Cy
2 25mM KCl 10mM HEPES(pH 7.5) protease inhibitors 1:200 Buffer B: 0.25mM sucrose in Buffer A Buffer N: 1.1mM sucrose in Buffer A PBS:137mM NaCl 2.7mM KCl 2mM KH2PO4 10mM Na2HPO4 pH=7.4 步骤(以下是我的实验笔记) 细胞量至少
I. NEBULIZATION of DNA 1. 0.5 - 5 ug DNA in TE (10mM/1mM), 25% glycerol, final volume 500 ul 2. nebulize for 90-100 sec at 5-10 psi (for a medium size of about 1.5kbp) 3. Precipitate with EtOH/Ammonium Acetate
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