So unfortunately I have low amounts of DNA used for my transformation. I still have the plates so was wondering if it's best to just pick a colony from that plate, grow an O/N. make a fresh plate then pick a colony for miniprep and sequencing?
That may be your best bet, if the DNA stock is lost.
However, E. coli plates aren't very stable - the colonies lose viability in a few weeks even at 4*C. Don't be too surprised if you don't recover viable bacteria.
It will be prudent to re-confirm the plasmid is what you're expecting by PCR, restriction digestion, and/or sequencing in case there was a mutation durring the long in vivo storage or durring recovery.
That may be your best bet, if the DNA stock is lost.
However, E. coli plates aren't very stable - the colonies lose viability in a few weeks even at 4*C. Don't be too surprised if you don't recover viable bacteria.
It will be prudent to re-confirm the plasmid is what you're expecting by PCR, restriction digestion, and/or sequencing in case there was a mutation durring the long in vivo storage or durring recovery.
Hey, you can just pick up different colonies and grew them separately in LB media for overnight in presence of antibiotics. For confirmation of the plasmid: restriction digestion or sequencing (optional).
If the plate is older than a month, There is maximum chance that you will not able to recover the bacteria, then try from your glycerol stock.
And also you can re-streak the colony into fresh agar plate.
Dear
You can do efficient experiment by using separated colonies. So you have to purified twice the colonies in the medium containing selective antibiotics. After purification the restriction digestion will followed. Be careful to don't use glycerol stock of bacterial culture stored.
Cordialy
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