From 2009.igem.org
Tuesday 4 August 2009
Results
Maria and Afton
- Overnight culture of I1352 plated on AMP:
- Sample taken from plate after PSB4C5 ligation attempt successfully grew expressing RFP. The color of the sample was pink
- Sample taken from plate after ligation with itself did not grow at all, and expressed no RFP
- There is a possibility the pSB4C5 backbone may be a bad part
- previous years it has not functioned properly
- Gel electrophoresis indicates DNA Gel Purification may be necessary
- Possible inefficiency of BioBrick enzymes may have come from repeated freezing and thawing
- They will be aliquoted into smaller amounts
- Gel Electrophoresis Results
From Left to Right: 100bp Ladder, Bioline Hyperladder, I1352(Digest), I1352(Miniprep), pSB4C5(Digest), pSB4C5(Miniprep)
Trentay 22:25, 4 August 2009 (UTC)
Tasks
Maria and Afton
- Run a Gel Electrophoresis
- Transform and grow up pSB1C3 with the death gene P1010
Trentay 22:11, 4 August 2009 (UTC)
Wetlab
- New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts
Name | Amount/Tube | # Tubes | Extra in Stock |
|
XbaI
| 10 µL | 15 | No |
|
SpeI
| 5 µL | 10 | No |
|
PstI
| 10 µL | 25 | Yes |
|
EcoRI-HF
| 10 µL | 25 | Yes |
|
BSA
| 5 µL | 8 | Yes |
|
T4 Ligase Buffer
| 10 µL | 25 | Yes |
|
T4 Ligase
| 10 µL | 8 | No |
|
- To show how many times a tube has been thawed and refrozen, a hash mark will be placed on the side of the tube, each time it is thawed as a reference for others to see
Trentay 15:24, 5 August 2009 (UTC)
Maria and Afton
- Gel Electrophoresis was run
- pSB4C5 and I1352 digested and undigested (right after Miniprep) were run on a gel and results showed excessive bands in the undigested DNA. DNA Gel Purification will be done on future Miniprepped DNA
- pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer.
- Transformation was done using DB 3.1
Name | Plate | Growth Observation |
|
pSB1C3
| CM | n/a |
|
(+) Control
| LB | n/a |
|
(+) Control zapped
| LB | n/a |
|
(-) Control
| CM | n/a |
|
Trentay 22:33, 4 August 2009 (UTC)
|