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SEPTEMBER: WEEK 2
September, 6th
Inoculum of I47, I48, I49 into 5 ml LB+Amp for TECAN experiment of the following day.
- Inoculum of I44-2 and I51-3 into 5 ml LB+Cm34 to evaluate MiniPrep yields.
- Inoculum of I44-2 and I51-3 into 5 ml LB+Cm12,5 to evaluate MiniPrep yields and for sequencing.
- Inoculum for MiniPrep and sequencing of:
- <partinfo>BBa_J23100</partinfo>_4C5
- <partinfo>BBa_J23101</partinfo>_4C5
- <partinfo>BBa_J23105</partinfo>_4C5
- <partinfo>BBa_J23106</partinfo>_4C5
- <partinfo>BBa_J23110</partinfo>_4C5
- <partinfo>BBa_J23114</partinfo>_4C5
- <partinfo>BBa_J23116</partinfo>_4C5
- <partinfo>BBa_J23118</partinfo>_4C5
- I7-A
- I8-5-D
Inoculum of cultures for TECAN test on self-inducible promoters:
- I14-1
- I15-1
- I16-1
- I17-1
- I18-1
- I19-1
- I14_4C5-2
- I15_4C5-2
- I16_4C5-1
- I17_4C5-1
- I18_4C5-1
- I19_4C5-1
- I6-3
- I3-1
- <partinfo>BBa_B0031</partinfo>
Glycerol stocks of MG43, MC42, MC43. Static TECAN test (absorbance, green fluorescence and red fluorescence) on 200 ul of these cultures.
Cultures were diluted in 4 ml final of LB in order to obtain the same lowest OD with the following formula: culture to remove and substitute with LB=FinalVolume-(lowestOD/misuredOD)*FinalVolume.
Centrifuge at 4000 rpm, 25°C for 5 min to pellet the culture, discard the surnatant and resuspend the pellet with 1 mL of M9. Static TECAN test (absorbance, green fluorescence and red fluorescence) on 200 ul of these cultures.
Liquid PCR was performed on MG43, MC42, MC43, MC1061 (negative control).
September, 7th
MiniPrep for:
| Culture | Quantifications (ng/ul)
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| <partinfo>BBa_J23100</partinfo>_4C5 | 11,8
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| <partinfo>BBa_J23101</partinfo>_4C5 | 39,7
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| <partinfo>BBa_J23105</partinfo>_4C5 | 25,4
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| <partinfo>BBa_J23106</partinfo>_4C5 | 29,7
|
| <partinfo>BBa_J23110</partinfo>_4C5 | 24,7
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| <partinfo>BBa_J23114</partinfo>_4C5 | 15,4
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| <partinfo>BBa_J23116</partinfo>_4C5 | 20,7
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| <partinfo>BBa_J23118</partinfo>_4C5 | 20,4
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| I7-A | 113,6
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| I8-5-D | 127,6
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| I44-2 (Cm 34) | 63,9
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| I51-3 (Cm 34) | 100,4
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| I44-2 (Cm 12,5) | 68,1
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| I51-3 (Cm 12,5) | 135,4
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| I47 | 74,8
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Cultures for TECAN test were diluted in the morning (h10:20) 1:100 (40ul in 4ml) and let grown at 37°C 220 rpm till they reach the desired OD.
September, 8th
Inoculum of:
- I31
- I32
- I34
- I35
- I36
- I37
- I40
- I41
- I20
into 5 ml LB + Amp;
already available:
- I21(E-S)
- I26(E-S)
- I0(E-X)
for second step of ligation of phasins/intein
September, 9th
Minipreps were quantified as follows:
- I31: ng/ul
- I32: ng/ul
- I34: ng/ul
- I35: ng/ul
- I36: ng/ul
- I37: ng/ul
- I40: ng/ul
- I41: ng/ul
- I20: ng/ul
September, 10th
September, 11th
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