Team:UNIPV-Pavia/Calendar/September/settimana2
From 2010.igem.org
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(→September, 10th) |
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Glycerol stocks of MG43, MC42, MC43. Static TECAN test (absorbance, green fluorescence and red fluorescence) on 200 ul of these cultures. | 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/ | + | 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/measuredOD)*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. | 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. | ||
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Trasformation of I42 and I43 in MC008 and MG008 strains, strain DB3.1 without DNA and pSB1C3 RFP in DB3.1 strain. | Trasformation of I42 and I43 in MC008 and MG008 strains, strain DB3.1 without DNA and pSB1C3 RFP in DB3.1 strain. | ||
MG42, MC43, MC42, MC43 were plated on LB+Cm34 plates instead DB3.1 on LB+Cm12.5 plates. | MG42, MC43, MC42, MC43 were plated on LB+Cm34 plates instead DB3.1 on LB+Cm12.5 plates. | ||
- | Integration protocol | + | Integration protocol first day were performed. |
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==September, 8th== | ==September, 8th== | ||
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+ | <font size=4>[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test8settembre|Tecan Test]]</font> | ||
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+ | ---- | ||
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Inoculum of: | Inoculum of: | ||
*I31 | *I31 | ||
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*I0(E-X) | *I0(E-X) | ||
for second step of ligation of phasins/intein | for second step of ligation of phasins/intein | ||
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<div align="right"><small>[[#indice|^top]]</small></div> | <div align="right"><small>[[#indice|^top]]</small></div> | ||
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---- | ---- | ||
- | MG42 and MG43 | + | MG42 and MG43 showed no colonies: probably the reason was a mistake on the choice of the MG008 strain tubes during the trasformations. |
<div align="right"><small>[[#indice|^top]]</small></div> | <div align="right"><small>[[#indice|^top]]</small></div> | ||
==September, 10th== | ==September, 10th== | ||
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+ | <font size=4>[[Team:UNIPV-Pavia/Material Methods/Measurements/Tecan/test10settembre_bis|Tecan Test]]</font> | ||
+ | |||
+ | ---- | ||
Screening digestion of I44-2 and I51-3. | Screening digestion of I44-2 and I51-3. | ||
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Gel run/cut for samples of previous day. Gel extraction showed the following quantifications: | Gel run/cut for samples of previous day. Gel extraction showed the following quantifications: | ||
- | *I31 (E-X): ng/ul | + | *I31 (E-X): 21.2 ng/ul |
- | *I32 (E-S): ng/ul | + | *I32 (E-S): 11.3 ng/ul |
- | *I35 (S-P): ng/ul | + | *I35 (S-P): 24.6 ng/ul |
- | *I36 (E-S): ng/ul | + | *I36 (E-S): 12.1 ng/ul |
- | *I37 (E-X): ng/ul | + | *I37 (E-X): 26 ng/ul |
- | *I37 (X-P): ng/ul | + | *I37 (X-P): 12.2 ng/ul |
- | *I40 (E-S): ng/ul | + | *I38 (E-S): 10.8 ng/ul |
- | *I41 (E-X): ng/ul | + | *I40 (E-S): 16.3 ng/ul |
- | *I20 (S-P): ng/ul | + | *I41 (E-X): 24.4 ng/ul |
+ | *I20 (S-P): 30.3 ng/ul | ||
Digestions were ligated in order to create: | Digestions were ligated in order to create: |
Latest revision as of 19:14, 27 October 2010
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