Let1 - that contains histidine and also to a plate that...

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Let’s say that we are interested in the E. coli genes that are involved in synthesis of histidine. To find insertion mutants that can not synthesize histidine (His ) we could screen amongst our collection of 2x10 4 random TTTTT insertions to find those that are His . The easiest way to do this would be to plate out the collection of insertions at a density of 200 colonies per plate (100 plates total). Each of these master plates would then be replica plated (first by transfer to a sterile piece of velvet) to a plate
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Unformatted text preview: that contains histidine and also to a plate that lacks histidine. His – insertion mutants would be identified as colonies that can not grow on the plates that lack histidine. Note that the same collection of random Tn5 insertions can be screened multiple times to find interesting mutations with different phenotypes. 3) Identify His – TTTTT insertion mutants by replica plating to find colonies that specifically can not grow on plates that don’t contain histidine....
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