From 2009.igem.org
Wet
GVP Cluster
Transporters
Below the PCR of 17/07, There seems to be a vague band at ~1150 in 1.
The band at ~1150 kb was cut out of the gel and used for pcr described below.
PCR1 gel
| Component | amount
|
| MasterMix NH4
| 40 uL
|
| HmtA Fw
| 1 uL
|
| HmtA mut1RC
| 1 uL
|
| PCR1 band
|
|
| Taq polymerase
| 2uL
|
| PCR1 program
| Temperature
| Time
|
| Denaturing
| 95°
| 5 min
|
|
| Start Cycles 30X
|
| Denaturing
| 95°
| 20 sec
|
| Annealing
| 55°
| 20 sec
|
| Elongation
| 72°
| 1.10 min
|
|
| End cycles
|
| Final elongation
| 72°
| 10 min
|
| Hold
| 4°
| Forever
|
Metal Accumulation
Vectors
positive control psB1AC3
VR-VF2
| Component | amount
|
| MasterMix NH4
| 21 uL
|
| F primer
| 1 uL
|
| R primer
| 1 uL
|
| psB1AC3 vector
| 0.5 uL
|
| Taq polymerase
| 1 uL
|
| Cup nr
| F primer
| R primer
|
| 1
| VF2 iGEMGr09
| VR iGEMGr09
|
| 2
| VF2 iGEMGr09
| VR iGEMGr08 Aug
|
| 3
| VF2 iGEMGr09
| VR iGEMGr June
|
| 4
| VF2 iGEMGr08
| VR iGEMGr09
|
| 5
| VF2 iGEMGr08
| VR iGEMGr08 Aug
|
| 6
| VF2 iGEMGr08
| VR iGEMGr June
|
| PCR1 program
| Temperature
| Time
|
| Denaturing
| 95°
| 5 min
|
|
| Start Cycles 30X
|
| Denaturing
| 95°
| 20 sec
|
| Annealing
| 55°
| 20 sec
|
| Elongation
| 72°
| 1.10 min
|
|
| End cycles
|
| Final elongation
| 72°
| 10 min
|
| Hold
| 4°
| Forever
|
Dry
|
"
