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From 2010.igem.org

July 8

1. Cloning issues: Did the double digestion of the terminator work?

• The EcoRI was old and might have failed
• SpeI not very efficient in EcoRI buffer, will try to incubate longer
• The ligation didn't work, but there are lots of ways to keep trying. This is a great way for people to come help during the week.

2.Copper precipitation assay

• Pick a concentration from the cell growth assay, like 1 mM Cu?
• Grow shaking, at time points spin out the cells and measure the absorbance
  • assaying for loss of copper

3. Does phsABC confer copper resistance? 96-well plate assay same as before, but with the phsABC plasmid

4. Cu resistance shipping in a day or two pRJ1004 (Cu efflux protein) (Jon) will email massive plasmid, no antibiotic resistance Get together next week to request several plasmids Tuesday morning at 10 am.

5. Email Kara about the mold (Jon)

6. Ways to get students involved (Dan email the group) Ask when they're available, delegate a task make sure someone is there to help them