From 2009.igem.org
Wet
LB-medium is badly needed!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
GVP Cluster
EM pictures
- → TODO Today we will try to get some Electon microscopy pictures of the gas vesicle producing bacteria from the ON cultures.
Orinigal pNL29 iptg-GVP and the biobricks ars-GVP and
- → TODO Choose colonies from plates for growth of E.coli TOP10 with GVP behind LAC/pBAD (+RBS) promoter in pSB1AC3
- → TODO Make glycerol stocks of pBad/araC, pBad/araC+RBS+GVP, and pLacI+RBS+GVP (both in pSB1A2 and pSB1AC3 plasmid)
- → TODO Test control of bouyancy in Saline solution (grow plates with GVP constructs)
- → TODO Order synthetic DNA for GVP
- → TODO Order primer for PstI site removal
- → TODO Test promoter strenght compared to BBa_J23101 promoter (Sven)
- → TODO Enter sequences of constructs to Sandbox
Colonies on Plates
Name
| Plasmid Used
| Antibiotics on Plasmid
| No. of Colonies
| Date
|
pSB2K3-BBa_J04450 (1,7C)
| pSB2K3
| Kanamycin
| 0
| 2/9
|
pSB2K3-BBa_J04450 (1,7C)(conc.)
| pSB2K3
| Kanamycin
| 4
| 2/9
|
pSB2K3-BBa_P1010 (1,7K)
| pSB2K3
| Kanamycin
| 0
| 2/9
|
pSB2K3-BBa_P1010 (1,7K)(conc.)
| pSB2K3
| Kanamycin
| 0
| 2/9
|
Negative Control
| MQ
| None
| 0
| 2/9
|
Positive Control
| J61002-J23101
| Ampicillin
| 0
| 2/9
|
- → The plates showed very low colony growth, and can be caused by the fact it is a low copy plasmid which can be induced by iptg to increase the copy number. Additional growth with IPTG should increase the amount of colonies and available plasmid.
- → The positive control failed due to growth of ampicillin resistance plasmids on kanamycin plates.
- → From the plate with four colonies 5mL LB-amp100 medium was inoculated, and additional plates were swiped with the used colonies to see if single colony growth on plate can occur.
O.n. precultures
Transporters
Metal Accumulation
- Send MBP-ArsR for sequencing again
- Send pLow-fMT for sequencing
- Send pLac-fMT for sequencing
Vectors
Dry