Team:UNIPV-Pavia/Calendar/September
From 2010.igem.org
Line 138: | Line 138: | ||
<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"> | ||
- | 88° Bio-Lab - | + | 88° 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"> | ||
- | 89° Bio-Lab - | + | 89° 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"> | ||
- | 90° Bio-Lab | + | 90° 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"> | ||
- | 91° Bio-Lab - | + | 91° Bio-Lab - Digestion of yesterday are repeted (because noone 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"> | ||
- | 92° Bio-Lab - | + | 92° 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, 25th|September, 25th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 25th|September, 25th]]</td><td align="left"> | ||
- | 93° Bio-Lab - | + | 93° Bio-Lab - |
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 26th|September, 26th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana4 #September, 26th|September, 26th]]</td><td align="left"> | ||
- | xx° Bio-Lab - | + | xx° 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"> | ||
- | 94° Bio-Lab - | + | 94° 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. | <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"> | ||
- | 95° Bio-Lab - | + | 95° 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 <parinfo>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"> | ||
- | 96° Bio-Lab | + | 96° Bio-Lab - Tecan Test was performed on prepared samples, after the usual protocol. |
+ | |||
+ | 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 <parinfo>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"> | ||
- | 97° Bio-Lab - | + | 97° 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> | ||
Revision as of 20:35, 3 October 2010
|
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
|