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: bit.ly/plasmid_prepping
more on cloning cells: blog form: bit.ly/dh5alpha ; RUclips: • DH5α strains and a PSA...  
more on expression cells: bit.ly/bl21_rare_codons ; RUclips: • Bacterial expression s...
more on molecular cloning: bit.ly/molecularcloningguide ; RUclips: • Molecular cloning over...
more on transformation: bit.ly/transformheatshock ; RUclips: • Bacterial transformati...
more on T7 RNAP: bit.ly/t7rnap
more on mini preps: bit.ly/minipreps ; RUclips: • Miniprep science - wha...  
more on glycerol solution-making: • Making a 80% glycerol ...
some tips for pipetting & working with viscous solutions: blog: bit.ly/pipetting_problems ; RUclips: • Pipetting viscous liqu...
random practical lab tips & tricks: bit.ly/lab_tricks_page
link to the plasmid I was talking about: www.addgene.org/174866/
helpful guide from addgene: www.addgene.org/recipient-ins...
more about all sorts of things: #365DaysOfScience All (with topics listed) 👉 bit.ly/2OllAB0 or search blog: thebumblingbiochemist.com                             
                               
#scicomm #biochemistry #molecularbiology #biology #sciencelife #science #realtimechem

[@Dominoes0]

This video is great (The diagrams, narration, show+tell) - it's more detailed and actionable than the videos and written guides provided by Addgene itself!

[@thebumblingbiochemist]

Thanks! Glad it was helpful!

[@Simon-si3ec]

Super helpful, thanks so much, just received my first plasmid and needed to visualize all these steps.

[@thebumblingbiochemist]

So great to hear! Best of luck!