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文献和实验Clone Genes From a Phage Library
). Its complexity is roughly 80,000 X an insert size of 4 to 8 kb = 320 Mbp, which is about 20-fold coverage of this yeast genome. The aim is to evenly plate out about 10,000 phage over about 10 plates. What you need in addition to the phage is: Plates: LB
To fix, 1/10 volume of 37 formaldehyde is added to the culture, which is shaken for a further 40 minutes at room temperature. Cells are then recovered by centrifugation at 1400 X g for 2 minutes, and washed (i.e. gently resuspended then recovered
Anti-GFP/BrdU In Situ Immunostaining
of 2 N HCl x30'. Wash 3x in PBS. Block in 1 mL 10% Horse serum, x30' at room temp. Wash 1x in PBS. Add 0.3 mL Biotinylated anti-BrdU (1:300, with 1% horse serum in PBS). Incubate at 4ºC o.n. or 2 hr. at room temp. Wash 3x in PBS.
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