Team:Groningen/Notebook/3 September 2009

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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
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

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