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本人小小研一学生一枚，求助求助。 2.4 Isolation of Phage Nucleic Acids 1)The phage filtrate having 8% polyethylene glycol (PEG) and 4% NaCl was centrifuged at 26,000 rpm for 4 h. 2) Dissolve the pellet in 100 μL autoclaved distilled water and sonicate it for 3 min. Centrifuge it again for 5 min at 14,000 rpm. 3)One microliter of DNaseI was added and incubated at 37°C for 30 min. After incubation,4 μL of 2.5 X SDS-EDTA dye (0.4% SDS, 30 mM EDTA, 0.25% bromophenol blue, 20% sucrose) was added to the tubes and kept at 80°C in Heat Block for 10 min. After heat treatment, the whole lysate was loaded onto 0.7% TAE agarose gel (Tris base, boric acid, EDTA, pH 8.3) containing 0.5 μg/mL of ethidium bromide. 4)The gel was run at a voltage of 90 V and the bands were visualized using a UV transilluminater. |
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