Team:UNIPV-Pavia/Calendar/September
From 2010.igem.org
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<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana1 #September, 4th|September, 4th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana1 #September, 4th|September, 4th]]</td><td align="left"> | ||
- | 74° Bio-Lab - | + | 74° Bio-Lab - Results of genomic integration of samples inoculated yesterday |
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana1 #September, 5th|September, 5th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana1 #September, 5th|September, 5th]]</td><td align="left"> | ||
- | 75° Bio-Lab - | + | 75° Bio-Lab - Plates were checked and new colonies were picked. |
</td></tr> | </td></tr> | ||
<tr><th align="center" width="12%" colspan="2"><b>SEPTEMBER: WEEK 2</b></th></tr> | <tr><th align="center" width="12%" colspan="2"><b>SEPTEMBER: WEEK 2</b></th></tr> | ||
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</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana2 #September, 7th|September, 7th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana2 #September, 7th|September, 7th]]</td><td align="left"> | ||
- | 77° Bio-Lab -MiniPreps of <partinfo> | + | 77° Bio-Lab -MiniPreps 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, I44-2, I51-3, I44-2, I51-3 and I47 were performed. |
Trasformation of I42 and I43. Integration protocol's first day were performed. | Trasformation of I42 and I43. Integration protocol's first day were performed. | ||
<br> | <br> | ||
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</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana2 #September, 9th|September, 9th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana2 #September, 9th|September, 9th]]</td><td align="left"> | ||
- | 79° Bio-Lab - | + | 79° Bio-Lab - Minipreps of I31, I32, I35, I36, I37, I38, I40, I41 and I20 were performed. |
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana2 #September, 10th|September, 10th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana2 #September, 10th|September, 10th]]</td><td align="left"> | ||
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Gel run/cut for samples of previous day (I31, I32, I35, I36, I37, I38, I40, I41 and I20). | Gel run/cut for samples of previous day (I31, I32, I35, I36, I37, I38, I40, I41 and I20). | ||
- | Digestions were ligated in order to create: I53, I54, I55, I56, I57, I58. | + | Digestions were ligated in order to create: I52, I53, I54, I55, I56, I57, I58. |
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana2 #September, 11th|September, 11th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana2 #September, 11th|September, 11th]]</td><td align="left"> | ||
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</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana3 #September, 17th|September, 17th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana3 #September, 17th|September, 17th]]</td><td align="left"> | ||
- | 86° Bio-Lab - | + | 86° Bio-Lab - Miniprep and Nanodrop quantification for: I58a, I58b, I58c, I59a, I59b, I59c, I60a, I60b, I60c, I61a, I61b and I61c. |
+ | |||
+ | Digestion (for screening) and gel extraction for: I58a/b/c, I59a/b/c, I60a/b/c. | ||
+ | |||
+ | Digestion (only screening) for I61a/b/c. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana3 #September, 18th|September, 18th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana3 #September, 18th|September, 18th]]</td><td align="left"> | ||
- | 87° Bio-Lab - | + | 87° Bio-Lab - Gel extraction and quantification of: I58, I59, I60. DNA was than stored at -20°C. |
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana3 #September, 19th|September, 19th]]</td><td align="left"> | ||
+ | 88° Bio-Lab - I55 was transformed again into E. coli DH5-alpha and plated on LB+Amp agar plate, that was let grow ON at 37°C. | ||
</td></tr> | </td></tr> | ||
<tr><th align="center" width="12%" colspan="2"><b>SEPTEMBER: WEEK 4</b></th></tr> | <tr><th align="center" width="12%" colspan="2"><b>SEPTEMBER: WEEK 4</b></th></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 20th|September, 20th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 20th|September, 20th]]</td><td align="left"> | ||
- | + | 89° Bio-Lab - Screening PCR on 6 colonies picked from I55 was performed. | |
+ | |||
+ | Inoculum of I52, I53, I55, I56 and I57 in LB+Amp for MiniPrep of the following day. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 21st|September, 21st]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 21st|September, 21st]]</td><td align="left"> | ||
- | + | 90° Bio-Lab - Miniprep and Nanodrop quantification for: I52, I53, I55, I56, I57; 3 hour digestion and gel run/cut were performed on this samples. | |
+ | Gel extraction was performed on I52, I56, I57, I61. | ||
+ | |||
+ | We already had digested DNA so we could perform new ON ligations: I62, I63, I64, I65, I66. | ||
+ | |||
+ | Tecan Test was performed on prepared samples, after a new protocol. | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 22nd|September, 22nd]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 22nd|September, 22nd]]</td><td align="left"> | ||
- | + | 91° Bio-Lab - Trasformation of ligations I62, I63, I64, I65 and I66 into E.coli TOP10. | |
+ | |||
+ | MiniPrep and quantification of: MyYeast1, MyYeast2, MyYeast3. Screening digestion was performed on this samples. | ||
+ | |||
+ | Glycerol stocks of MyYeast1, MyYeast2, MyYeast3 was performed. | ||
+ | |||
+ | Inoculum in LB+Amp+Cm12,5 from glycerol stocks and colony for antibiotic resistance testing of I47_S1, I48_S1, I49_S1 and I13522. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 23rd|September, 23rd]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 23rd|September, 23rd]]</td><td align="left"> | ||
- | + | 92° Bio-Lab - Digestion of yesterday are repeted (because none of them were postitive). This time all samples seems to be positive. | |
+ | |||
+ | Inoculum for three colonies from I62, I63, I64, I65 agar plates into 5ml LB+Amp. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 24th|September, 24th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 24th|September, 24th]]</td><td align="left"> | ||
- | + | 93° Bio-Lab - Glycerol stocks was performed for I62, I63, I64, I65, I66. | |
+ | |||
+ | MiniPrep and Nanodrop quantification : I62-1, I62-2, I62-3, I63-1, I63-2, I63-3, I64-1, I64-2, I64-3, I65-1, I65-2, I65-3, I66-1, I66-2 and I66-3. | ||
+ | Digestion for screening and gel run of samples were performed. | ||
+ | |||
+ | Dilution 1:50 of I47, I48, I49, GFP in LB+Amp+Cm12,5 and S1 in LB+Cm12,5. | ||
</td></tr> | </td></tr> | ||
- | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, | + | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 26th|September, 26th]]</td><td align="left"> |
- | + | 94° Bio-Lab - Inoculum of <partinfo>BBa_K173000</partinfo>, I47, I48, I49, <partinfo>BBa_B0031</partinfo> into 3 ml LB+Amp for tomorrow TECAN test. | |
+ | |||
+ | Pick of six colonies from I66 agar plate (I66-A/B/C/D/E/F) and inoculum into 3 ml LB+Amp for screening thorough E-P digest. | ||
</td></tr> | </td></tr> | ||
<tr><th align="center" width="12%" colspan="2"><b>SEPTEMBER: WEEK 5</b></th></tr> | <tr><th align="center" width="12%" colspan="2"><b>SEPTEMBER: WEEK 5</b></th></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana5 #September, 27th|September, 27th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana5 #September, 27th|September, 27th]]</td><td align="left"> | ||
- | + | 95° Bio-Lab - MiniPrep and E-P digest for I62-a, I62-b, I62-c, I62-d, I62-e, I62-f. | |
+ | |||
+ | DNA PCR for MyYeast 1-2-3 | ||
+ | |||
+ | Glycerol stock was performed for I62-e and stored it at -80°C. | ||
+ | |||
+ | Protein electrophoresis for phasins. | ||
+ | <br>14° Meeting - Updating about Bio-Lab activity. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana5 #September, 28th|September, 28th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana5 #September, 28th|September, 28th]]</td><td align="left"> | ||
- | + | 96° Bio-Lab - Tecan Test was performed on prepared samples, after the usual protocol. | |
+ | |||
+ | Miniprep and quantification for: I1, I4, I53, <partinfo>BBa_J04450</partinfo>-1, <partinfo>BBa_J04450</partinfo>-2. | ||
+ | Digestion of I1, I4, I53, <partinfo>BBa_J04450</partinfo>-1, <partinfo>BBa_J04450</partinfo>-2, MyYeast-3 (DNA already available) and <partinfo>pSB4C5</partinfo>. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana5 #September, 29th|September, 29th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana5 #September, 29th|September, 29th]]</td><td align="left"> | ||
- | + | 97° Bio-Lab - Colony PCR for 6 newly picked colonies. Gel run for all digested parts and cut/gel purification for:4C5 , MyYeast-3, I1, I4, I53, <partinfo>BBa_J04450</partinfo> and <partinfo>pSB4C5</partinfo>. | |
+ | |||
+ | Ligation of:I70=MyYeast 3+I4, I71=MyYeast 3+I1, I72=BBa_J04450+pSB4C5 and I73= I38+I53. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana5 #September, 30th|September, 30th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana5 #September, 30th|September, 30th]]</td><td align="left"> | ||
- | + | 98° Bio-Lab - Tecan Test was performed on prepared samples, after the usual protocol. | |
+ | |||
+ | Transformation of I73 (1ul) into E. coli TOP10. It was plated on LB+Amp 100 ug/ml agar plate. | ||
+ | |||
+ | Trasformation of: I70 in TOP10 and plated on LB+Amp, I71 in TOP10 and plated on LB+Amp and I72 in TOP10 and plated on LB+Cm 12,5. All plates were incubated at 37°C ON. | ||
+ | |||
+ | Miniprep and quantification of I20 was performed. Gel run/cut/extraction/quantification for I20. We repeated I55 ligation (I38+I20). | ||
+ | |||
+ | Inoculum into 5ml LB+Amp of: I31, I35, I37,I57,I60, INTEIN, 3x BBa_J04450 (to take pSB1C3, so pSB1C3 from now on). Cultures were let grow ON at 37°C, 220 rpm. | ||
+ | |||
+ | Inoculum of E. coli TOP10 in 1,5 ml LB, grown ON at 37°C 220 rpm. Tomorrow we will prepare competent cells!! | ||
+ | |||
+ | Inoculum of: S1 in 500ul M9+Cm 12,5; I47-S1 in 500ul M9+Amp+Cm 12,5; GFP-S1 in 500ul M9+Amp+Cm 12,5; RBS32 in 500ul M9+Amp. Cultures were grown ON at 37°C 220 rpm. | ||
+ | |||
+ | Tomorrow we will test the ability of these strains to produce GFP and to lyse when HSL is added. | ||
</td></tr> | </td></tr> | ||
Latest revision as of 19:16, 27 October 2010
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