Team:Hawaii/Notebook/2008-07-22

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Things we did today

Wetlab work

Alkaline Lysis Mini-prep

Margaret
  • Plasmid prep of pSB1A2, pSB1A3, and pSB3K3: 3 1.5 mL preps of the first two, and 10 of the third because it has a lower copy number. Used the protocol from Dr. Callahan's lab (http://packrat.stjohn.hawaii.edu/prestinglab/wiki/Protocols).
  • I had to skip the phenol/chloroform step, so after adding Rnase A and incubating for 20 minutes, I spun down again, collected the supernatant and then precipitated with isopropanol.
Grace
  • nir, pilA, slr-1, slr-2, GFP fusion

Glycerol Stock

Margaret
  • glycerol stock of pSB1A2, pSB1A3, and pSB3K3. These DB3.1 strains containing these plasmids can be found in the -80C with the other plasmids in E. coli.
Grace
  • nir, pilA, slr-1, slr-2, GFP fusion, BB-pRL1383a, BBa_E0026

pRL1383a insert Clean-up

Margaret
  • in preparation for constructing the biobrick form of pRL1383a, cleaned the PCR products: mob, rep, oriV, and the aadA region using the "Isopropanol Precipitation for PCR purification" protocol from openwetware.org
  • Cleaned the oriT construct using the Qiagen Minelute Gel Purification kit.

Made 80% glycerol stock solution

Grace

Organized plasmid preps

Grace
  • Determined DNA concentrations of all plasmid preps to date
  • Relabeled all tubes w/ plasmid (or BioBrick part), DNA concentration, date of prep, and initials of person who prepped
  • Posted more detailed plasmid prep information on private website

Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


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