User:Wilfred J. Poppinga/Notebook/Wilfreds Project/2009/08/08

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Weekend stuff iGEM Main project page
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Miscellaneous

  • Check plates Nienke
  • Check plates Frans

Comments

  • Frans his plates were barely grown on the bench, put them to 37 °C for 5 hours.

Results

Nienke's plates MymT
Colonies Name Plasmid Date of plating Antibiotics
226
MymT Conc pSB68(MymT) 7august2009 LB-Amp
∞/10
pSB1AC3-lowP 7august2009 LB-Cam150
12
MymT p4B68(MymT) 7august2009 LB-Amp
0
–-
pSB1AC3 7august2009 LB-Cam150
(+)
pSB1AC3-lowProm 7august2009 LB-Cam150
>400
(MymT Conc) pGB68(MymT) conc. 7august2009 LB-Amp100
>400
(MymT) pGB68(MymT) 7august2009 LB-Amp
4
–-
pSB1AC3 7august2009 LB-Cam150

Everything was written as I read it, no guarantees.

Frans' plates
Colonies Name Plasmid Date of plating Antibiotics
0
K3 pBad 2:3 pSB3K3 7august2009 50 μg/mL kan
0
K3 pBad 2:3 pSB3K3 7august2009 50 μg/mL kan
0
K3 pBad 2:4 pSB3K3 7august2009 50 μg/mL kan
3
K3 pBad 2:4 pSB3K3 7august2009 50 μg/mL kan
~150
AC3 p4od 1:3 pSB1AC3 7august2009 LB AMP
~30
AC3 pBad 1:3 pSB1AC3 7august2009 LB AMP
1
AC3 Lac 1:4 pSB1AC3 7august2009 LB AMP
3
AC3 Lac 1:4 pSB1AC3 7august2009 LB AMP

After 5 h at 37 °C (and ON on the bench) Codes are written as they are read, not as they are interpreted. Plates with no growth were put back on the bench.

ALL PLATES WITH GROWTH, Nienke & Frans, were put @ 4 °C



Literature stuff Pharmaceutical biology Main project page
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Discussion

  • Dyson et al.; Production of soluble mammalian proteins in Escherichia coli: identification of protein features that correlate with successful expression, BMC Biotechnology 2004, 4:32 doi:10.1186/1472-6750-4-32
    • Trx and MBP are best fusion proteins, recommended is to truncate proteins >50 kDa
    • They barely express soluble cell surface receptors



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