Team:Yale/MeetingNotes

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==July 8==
==July 8==
-
1. Cloning issues:
+
The phsABC plasmid worked!  We have metal precipitation!  Woooo!
-
Did the double digestion of the terminator work?
+
-
* The EcoRI was old and might have failed
+
-
SpeI not very efficient in EcoRI buffer, run longer
+
-
ligation didn't work, keep trying
+
 +
1. Cloning issues
 +
*Did the double digestion of the terminator work?
 +
** The EcoRI was old and might have failed, check each enzyme on a gel
 +
*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
+
2. Copper precipitation assay
-
1 mM Cu?
+
*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
+
*Grow shaking, at time points spin out the cells and measure the absorbance
-
assaying for loss of copper
+
**this is assaying for loss of copper
3. Does phsABC confer copper resistance?
3. Does phsABC confer copper resistance?
-
96-well plate assay
+
*96-well plate assay
-
same as before, but with the phsABC plasmid
+
*same as before, but with the phsABC plasmid
-
 
+
**Since Leah has this assay dialed-in, this is a great job for a part-time helper to run and analyze!
4.  Cu resistance  
4.  Cu resistance  
-
shipping in a day or two
+
* will be shipping in a day or two
-
pRJ1004 (Cu efflux protein) (Jon) will email
+
**pRJ1004 (Cu efflux protein) (Jon will email to follow up)
-
massive plasmid, no antibiotic resistance
+
***massive plasmid, no antibiotic resistance
-
Get together next week to request several plasmids
+
**Get together next week to request several plasmids (Tuesday morning at 10 am).
-
Tuesday morning at 10 am.
+
5. Email Kara about the mold (Jon)
5. Email Kara about the mold (Jon)
6. Ways to get students involved (Dan email the group)
6. Ways to get students involved (Dan email the group)
-
Ask when they're available, delegate a task
+
*Ask when they're available, delegate a task
-
make sure someone is there to help them
+
*make sure someone is there to help them
 +
 
 +
==July 15==

Latest revision as of 21:24, 8 July 2010

July 8

The phsABC plasmid worked! We have metal precipitation! Woooo!

1. Cloning issues

  • Did the double digestion of the terminator work?
    • The EcoRI was old and might have failed, check each enzyme on a gel
  • 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
    • this is assaying for loss of copper


3. Does phsABC confer copper resistance?

  • 96-well plate assay
  • same as before, but with the phsABC plasmid
    • Since Leah has this assay dialed-in, this is a great job for a part-time helper to run and analyze!

4. Cu resistance

  • will be shipping in a day or two
    • pRJ1004 (Cu efflux protein) (Jon will email to follow up)
      • 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

July 15