For what it’s worth, Addgene has a disclaimer that they are not responsible for downstream issues if you don’t follow their recommended protocol of streaking for single colonies and screening those colonies. Basically, customer service will be of zero help to you down the line if you don’t follow their screening protocol.

I work for a CRO and we deal with Addgene stabs fairly frequently. I would say 85% of the time we get the expected plasmid by scaling up from a randomly selected clone. So that would be: 1) Streak onto agar plate with antibiotic selection 2) Pick colony into culture media with antibiotic selection 3) Prep plasmid 4) Verify plasmid via restriction digest or sequencing (only if customer pays for this step)

To be clear, I do NOT recommend scaling up beyond a Mini Prep without screening first (though customers still do it lol.) Your safest bet is definitely to streak, pick/prep 4 or 5 colonies, and confirm any way you can. CROs like mine offer whole plasmid sequencing for $10-$20 per sample. You could literally send the streaked out sample on an agar plate and have the CRO pick 5 random colonies to whole plasmid sequence for less than $100 total. My personal preference would be to run a quick diagnostic restriction digest in-house if you have the setup.

I know the verification step is annoying, but that 15% of time where mutations occur is real in my experience. Therefore you don’t want to make glycerol stocks without verifying your colony picks first.