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Team:Groningen/Notebook/3 September 2009
From 2009.igem.org
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===GVP Cluster=== | ===GVP Cluster=== | ||
''' EM pictures ''' | ''' EM pictures ''' | ||
| - | Today we will try to get some Electon microscopy pictures of the gas vesicle producing bacteria from the ON cultures. | + | |
| + | :→ {{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 | 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''' | ||
| + | |||
| + | {|cellpadding="2" cellspacing="1" border="4" | ||
| + | |'''Name''' | ||
| + | |'''Plasmid Used''' | ||
| + | |'''Antibiotics on Plasmid''' | ||
| + | |'''No. of Colonies''' | ||
| + | |'''Date''' | ||
| + | |- | ||
| + | |[http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_J04450 BBa_J04450] (1,7C) | ||
| + | |pSB2K3 | ||
| + | |Kanamycin | ||
| + | |0 | ||
| + | |2/9 | ||
| + | |- | ||
| + | |[http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_J04450 BBa_J04450] (1,7C)(conc.) | ||
| + | |pSB2K3 | ||
| + | |Kanamycin | ||
| + | |4 | ||
| + | |2/9 | ||
| + | |- | ||
| + | |[http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_P1010 BBa_P1010] (1,7K) | ||
| + | |pSB2K3 | ||
| + | |Kanamycin | ||
| + | |0 | ||
| + | |2/9 | ||
| + | |- | ||
| + | |[http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_P1010 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-amp<sub>100</sub> 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=== | ===Transporters=== | ||
Revision as of 06:49, 3 September 2009
[http://2009.igem.org/Team:Groningen http://2009.igem.org/wiki/images/f/f1/Igemhomelogo.png]
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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 |
| [http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_J04450 BBa_J04450] (1,7C) | pSB2K3 | Kanamycin | 0 | 2/9 |
| [http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_J04450 BBa_J04450] (1,7C)(conc.) | pSB2K3 | Kanamycin | 4 | 2/9 |
| [http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_P1010 BBa_P1010] (1,7K) | pSB2K3 | Kanamycin | 0 | 2/9 |
| [http://partsregistry.org/Part:pSB2K3 pSB2K3]-[http://partsregistry.org/Part:BBa_P1010 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
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