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文献和实验that these four collections will each have gaps, due to (a) some genes being essential, (b) failure to obtain transformants that passed all 5 PCR confirmation tests. Central Storage: We will be storing 4 sets of A isolates in 2-ml tubes. For essential genes
Sequencing Reaction Cleanup in 384 Well Sequencing Reaction Plates
1.Take the 384 well viper plate from the thermocycler. Remove the Silicone Sealing Mat (Axygen Scientific: AM-384-PCR-RD) and place face down on a paper towel and pat to absorb any liquid that may be on it. Add 30 μl 95% EtOH/0.12 M NaOAc
, then centrifuged and resuspended in saline or live P. aeruginosa cells grown to mid log phase, then centrifuged and resuspended in saline. (For either method resuspend the bacteria so the concentration is 5 X 108 bacteria/ml.) In both methods the cell
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