Team:Yale/MeetingNotes
From 2010.igem.org
(Difference between revisions)
Dspakowicz (Talk | contribs) (→July 8) |
Dspakowicz (Talk | contribs) (→July 8) |
||
Line 4: | Line 4: | ||
Did the double digestion of the terminator work? | Did the double digestion of the terminator work? | ||
* The EcoRI was old and might have failed | * The EcoRI was old and might have failed | ||
- | SpeI not very efficient in EcoRI buffer, | + | *SpeI not very efficient in EcoRI buffer, will try to incubate longer |
- | ligation didn't work, keep trying | + | * 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 | 2.Copper precipitation assay | ||
- | Pick a concentration from the cell growth assay | + | *Pick a concentration from the cell growth assay, like 1 mM Cu? |
- | 1 mM Cu? | + | *Grow shaking, at time points spin out the cells and measure the absorbance |
- | Grow shaking, at time points spin out the cells and measure the absorbance | + | **assaying for loss of copper |
- | + | ||
Revision as of 21:10, 8 July 2010
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