What to do when a plasmid comes for you: agar stab to plate to glycerol stock, miniprep & sequencing

I ordered a bunch of plasmids, and many of them come as agar stabs, where they've taken bacteria with the plasmid and stabbed it into a tube of agar. So it's like a Petri dish in a tube. My gameplan:
streak a plate
isolate a colony
grow a 5mL overnight culture (LB + appropriate antibiotic)
make a glycerol stock
-- take 500μL of that culture & add to 500μL sterile glycerol; invert to mix & store in -80°C freezer - should be good for years if you avoid freeze-thaws
miniprep the rest of the overnight culture to isolate the plasmid
send the plasmid for sequencing
if the sequence checks out, do whatever the heck you got it for! (subclone into a different vector, transform into expression cells, etc.)

blog form: https://bit.ly/plasmid_prepping

more on cloning cells: blog form: https://bit.ly/dh5alpha ; YouTube:    • DH5α strains and a PSA to sequence your pl...    

more on expression cells: https://bit.ly/bl21_rare_codons ; YouTube:    • Bacterial expression strains - esp BL21 st...  

more on molecular cloning: http://bit.ly/molecularcloningguide ; YouTube:    • Molecular cloning overview - techniques & ...  

more on transformation: http://bit.ly/transformheatshock ; YouTube:    • Bacterial transformation - chemically comp...  

more on T7 RNAP: http://bit.ly/t7rnap

more on mini preps: http://bit.ly/minipreps ; YouTube:    • Miniprep science - what goes on in each st...    

more on glycerol solution-making:    • Making a 80% glycerol stock solution - a s...  

some tips for pipetting & working with viscous solutions: blog: https://bit.ly/pipetting_problems ; YouTube:    • Pipetting viscous liquids - reverse pipett...  

random practical lab tips & tricks: https://bit.ly/lab_tricks_page 

link to the plasmid I was talking about: https://www.addgene.org/174866/

helpful guide from addgene: https://www.addgene.org/recipient-ins...

more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 http://bit.ly/2OllAB0 or search blog: http://thebumblingbiochemist.com                             
                               
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