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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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#Use the stock solution to transform a recA, endA E.coli strain like TOP10, DH5alpha, JM109, or equivalent. | #Use the stock solution to transform a recA, endA E.coli strain like TOP10, DH5alpha, JM109, or equivalent. | ||
#Select transformants on [[Team:Mississippi State/Recipes|Low Salt LB plates]] containing 100ug/ml blasticidin. | #Select transformants on [[Team:Mississippi State/Recipes|Low Salt LB plates]] containing 100ug/ml blasticidin. | ||
| - | #Prepare a glycerol stock from each transformant containing plasmid for [[Team:Mississippi State/ | + | #Prepare a glycerol stock from each transformant containing plasmid for [[Team:Mississippi State/Recipes|long-term storage]]. |
Latest revision as of 16:12, 16 June 2008
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:
- Resuspend each vector in 20ul sterile water to prepare a 1ug/ul stock solution. Store the stock solution at -20C
- Use the stock solution to transform a recA, endA E.coli strain like TOP10, DH5alpha, JM109, or equivalent.
- Select transformants on Low Salt LB plates containing 100ug/ml blasticidin.
- Prepare a glycerol stock from each transformant containing plasmid for long-term storage.
