My culture with positive bacteria in it is unusable, and the plates with positive insert bacteria (E. coli) are stored @ 4 deg. cel. and I can't get them to regrow in an overnight culture. I do, however, have the miniprep product I produced from those cultures. My question is can I use this to transfect a new batch of E. coli?
yes, i have used the with insert miniprep to heat shock fresh batch ecoli and it worked.
Yes. Easily.
Use very little plasmid (1-10 ng is plenty for a pure plasmid)
We heat shock to transform chemically competent Dh5 alpha E. Coli.
Yes you can. Using pure plasmid, you may not even need to heat shock if the cells are sufficiently competent, a big time saver.
Dear Kolitha, the presence of RNA does not in my experience affect transformation efficiency. I regularly transform bacteria with plasmid prepared using a boiling miniprep, which contains significant amounts of contaminating RNA.
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