Blue colonies and a few White colonies are chosen to be cultured for future research.
The substrates' unequal distribution could be the cause. It's possible that certain colonies formed earlier and produced more enzyme. You might have some mixed colonies or scarce substrate if colonies are very dense. An in-frame insert may also result in light blue.
White colonies, where X-gal is not hydrolyzed, indicate the presence of an insert in lacZ that prevents the formation of an active -galactosidase, whereas blue colonies suggest they may contain a vector with an uninterrupted lacZ (therefore no insert).
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