I’ve been culturing what should be a pure strain of E. coli for a plasmid prep, but my last two plates show two distinct colony morphologies. I’m worried a contaminant has slipped in, maybe during my last transformation, and now I’m not sure which colonies to pick for my miniprep.
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What colonies should I pick for a plasmid miniprep with mixed morphologies?
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Samuel.M
Junior Member
OliviaYM
Junior Member
Two colony types showed up and it set off the alarm bells. In my experience that can mean a real contamination, but it can also be local microenvironment effects on the plate or subpopulations with different copy numbers. Hard to tell from colony shape alone.
Savannah_T
Junior Member
I’ve been there. I paused, noted the two looks, and looped in someone else before committing to any picks. In the end I leaned on isolation fundamentals rather than guessing based on morphology.
SophiaL
Junior Member
Sometimes it’s not a bad actor at all—plasmid burden or copy number variation can make colonies look different. It felt like chasing my tail for a while.
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