Team:Mississippi State/Maintenance of Plasmids

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(New page: =Maintenance of Plasmids= *The pPIC6alpha vector contains the blasticidin resistance gene to allow selection of the plasmid using blasticidin. To propogate and maintain the pPIC6alpha pl...)
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Maintenance of Plasmids

  • The pPIC6alpha vector contains the blasticidin resistance gene to allow selection of the plasmid using blasticidin. To propogate and maintain the pPIC6alpha plasmid, use the following procedure:
  1. Resuspend each vector in 20ul sterile water to prepare a 1ug/ul stock solution. Store the stock solution at -20C
  2. Use the stock solution to transform a recA, endA E.coli strain like TOP10, DH5alpha, JM109, or equivalent.
  3. Select transformants on Low Salt LB plates containing 100ug/ml blasticidin.
  4. Prepare a glycerol stock from each transformant containing plasmid for long-term storage.
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