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 for the psB1AC3 vector done with both new and last years primers.
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
|
Primers
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
|
PCR program
VR VF2 program
| Temperature
| Time
|
Denaturing
| 95°
| 5 min
|
| Start Cycles 30X
|
Denaturing
| 95°
| 20 sec
|
Annealing
| 55°
| 20 sec
|
Elongation
| 72°
| 20 sec
|
| End cycles
|
Final elongation
| 72°
| 10 min
|
Hold
| 4°
| Forever
|
The gel shown below shows bands at about 1000 which is not what would be expected. Band of 315 were expected.
Dry
|