IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/06/27

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Plasmid Mini-Preps

Mini-Preps were preformed on the 5 parts.

Procedure:

  • 1.5 mL of the transformed bacterial cultures were added to five 1.5 ml micro-centrifuge tubes. This was spun down, the supernantant removed, and repeated.
  • 100 μL of 7x lysis buffer was added to the pellets and re-suspended
  • Immediately after, 350 μL of cold neutralization buffer was added and the tube was mixed until all the blue was removed.
  • The tubes were centrifuged at 13.3 x g for 3 minutes
  • The supernantant was removed and each sample was placed into a Zymo-Spin column.
  • The column was placed in a collection tube and centrifuged at the same speed for 15 seconds
  • The flow through was discarded
  • 200 μL of Endo-Wash buffer was added to each column, then centrifuged for 30 seconds, then 400 μL of Zyppy wash buffer was added to each column and they were centrifuged for 1 min
  • the columns were transferred to clean 1.5 ml micro-centrifuge tubes and 30 μL of Zyppy elution buffer was added to the column and it was centrifuged for 30 seconds. Each sample had 5 columns, and these were all combined into 5 tubes, one for each sample.


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