"Back up" your clones! What to do after you confirm your plasmid sequence

Описание: After you confirm your clone sequence:

So you don’t get confused in the future and use the wrong tubes:
Discard* all the “bad” ones (plasmids with incorrect sequences and/or poor quality) ASAP
* Place (warningly-labeled) in a “to be discarded” box or other location for samples you plan to toss, but want to keep temporarily just in case
As long as you got good one(s): discard* the plate with potentially more “bad” ones
More thoroughly label good tube(s)

Next steps:
Transform into expression cells (e.g. BL21(DE3)) if desired

Saving for the future:
Re-transform into cloning cells, plate
Pick a colony from the new plate & grow an overnight culture
From the overnight culture prepare glycerol stocks for long-term storage, do an additional miniprep if you need more plasmid

And do be sure to sequence any plasmid you get! Whole plasmid sequencing technology now makes this cheap ($15 from Plasmidsaurus (not a sponsored post or anything, just what I use), quick (overnight results in my case), and easy (no primers needed)!

More on plasmid sequencing: http://bit.ly/sequenceclones &    • Whole plasmid sequencing with nanopore tec...  

More on glycerol stock preparation: https://bit.ly/plasmid_prepping

More on cloning vs. expression cells:    • Cloning cells (e.g. DH5α) vs. expression c...  
cloning cells: https://bit.ly/dh5alpha
expression cells: https://bit.ly/bl21_rare_codons

More on molecular cloning: http://bit.ly/molecularcloningguide

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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