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How not to do a miniprep

  • AM miniprep failed when tubes caps came off in initial cell-pelleting stage, splashing and mixing samples.
  • Re-inoculated 5 mL liquid cultures of LE392 transformants (B0015, J23114, R0011, C0012), 2 apiece in Amp LB and grew all day for PM miniprep
  • When liquid cultures reached OD of approximately 0.6, mixed 0.5 mL of cell solution from each culture with 0.5 mL of filter-sterilized 50% glycerol solution for long-term storage. Because didn't have immediate access to liquid nitrogen, left samples aside at 4 C to be flash-frozen the next morning.
  • Miniprepped the result in the evening according to the standard miniprep protocol, and left eluted DNA at 4 C.

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