Team:UNIPV-Pavia/Calendar/June/settimana4
From 2010.igem.org
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+ | <table class="menu" border="0" width="100%"> | ||
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+ | <td align="center" style="padding:0; height:20px"> | ||
+ | [[Team:UNIPV-Pavia/Calendar/June/settimana2|Week 2]] | ||
+ | </td> | ||
+ | <td align="center"> | ||
+ | [[Team:UNIPV-Pavia/Calendar/June/settimana3|Week 3]] | ||
+ | </td> | ||
+ | <td align="center"> | ||
+ | [[Team:UNIPV-Pavia/Calendar/June/settimana4|Week 4]] | ||
+ | </td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | <br> | ||
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+ | <html><p align="center"><font size="4"><b>JUNE: WEEK 4</b></font></p></html><hr><br> | ||
+ | <html><a name="indice"/></html> | ||
==June, 21st== | ==June, 21st== | ||
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Other colonies resulted positives at the screening but were NOT sequenced and are stored in the ''iGEM 2010 cemetery'' box. These clones are: I1-1, I2-2, I4-1 and I6-3. | Other colonies resulted positives at the screening but were NOT sequenced and are stored in the ''iGEM 2010 cemetery'' box. These clones are: I1-1, I2-2, I4-1 and I6-3. | ||
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+ | <div align="right"><small>[[#indice|^top]]</small></div> | ||
==June, 22nd== | ==June, 22nd== | ||
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Inoculum of I6, <partinfo>BBa_J23118</partinfo>, <partinfo>BBa_J23110</partinfo>, <partinfo>BBa_J23114</partinfo>, <partinfo>BBa_J23116</partinfo> from glycerol stock in 5ml LB+Amp. Cultures were grown ON at 37°C 220rpm. | Inoculum of I6, <partinfo>BBa_J23118</partinfo>, <partinfo>BBa_J23110</partinfo>, <partinfo>BBa_J23114</partinfo>, <partinfo>BBa_J23116</partinfo> from glycerol stock in 5ml LB+Amp. Cultures were grown ON at 37°C 220rpm. | ||
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+ | <div align="right"><small>[[#indice|^top]]</small></div> | ||
==June, 23rd== | ==June, 23rd== | ||
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Today we received 2 stabs from iGEM HQ for <partinfo>BBa_K208001</partinfo> (one for each registry location of this part), the Silver-fusion compatible BioBrick part that codes for phasin. This part is in a pSB3K3 plasmid. Bacteria were streaked on LB+Kan (50 ug/ml) agar plates. Cultures were named PhaP-1 and PhaP-2. Plates were incubated ON at 37°C. | Today we received 2 stabs from iGEM HQ for <partinfo>BBa_K208001</partinfo> (one for each registry location of this part), the Silver-fusion compatible BioBrick part that codes for phasin. This part is in a pSB3K3 plasmid. Bacteria were streaked on LB+Kan (50 ug/ml) agar plates. Cultures were named PhaP-1 and PhaP-2. Plates were incubated ON at 37°C. | ||
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+ | <div align="right"><small>[[#indice|^top]]</small></div> | ||
==June, 24th== | ==June, 24th== | ||
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Ligations were heated at 65°C for 5 minutes to inactivate ligase ad then tranformed in E. coli TOP10 home made competent cells. Plates were incubated ON at 37°C (we decided to incubate plates 5 hours longer than usual, because cell growth was already slower for I6). | Ligations were heated at 65°C for 5 minutes to inactivate ligase ad then tranformed in E. coli TOP10 home made competent cells. Plates were incubated ON at 37°C (we decided to incubate plates 5 hours longer than usual, because cell growth was already slower for I6). | ||
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+ | <div align="right"><small>[[#indice|^top]]</small></div> | ||
==June, 25th== | ==June, 25th== | ||
- | All plates showed colonies after 19 | + | All plates showed colonies after 19-hour growth at 37°C!! |
*I7: small green colonies with satellite colonies were observed. Negative colonies are present and can be easily distinguished, because they are red. | *I7: small green colonies with satellite colonies were observed. Negative colonies are present and can be easily distinguished, because they are red. | ||
*I8: green colonies were observed, few negative colonies are pink. | *I8: green colonies were observed, few negative colonies are pink. | ||
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These results are encouraging, because they are consistent with our expectations: ligations provide a high metabolic burden to the cell, so growth is slower. ''Positive'' green colonies are smaller and not surrounded by satellite colonies. ''Negative'' colonies are bigger and surrounded by satellite colonies, confirming that they had a faster growth than our ligations! | These results are encouraging, because they are consistent with our expectations: ligations provide a high metabolic burden to the cell, so growth is slower. ''Positive'' green colonies are smaller and not surrounded by satellite colonies. ''Negative'' colonies are bigger and surrounded by satellite colonies, confirming that they had a faster growth than our ligations! | ||
- | + | Two colonies from each plate were picked and incubated in 1ml LB+Amp. | |
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- | Cultures were grown for 6 | + | Cultures were grown for 6 hours at 37°C 220 rpm, then glycerol stocks were prepared for each culture. Glycerol stocks are stored at -80°C in ''iGEM 2010 ligations'' box. |
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Digestion was performed for 2 hours at 37°C. DNA was then gel run for screening. Both cultures were positive! Sequencing will be performed next week! | Digestion was performed for 2 hours at 37°C. DNA was then gel run for screening. Both cultures were positive! Sequencing will be performed next week! | ||
- | [[Image:Unipv_screening_PhaP_1_2.jpg| | + | [[Image:Unipv_screening_PhaP_1_2.jpg|200px|thumb|center|Screening of PhaP-1 and PhaP-2]] |
+ | <div align="right"><small>[[#indice|^top]]</small></div> | ||
+ | <!-- table previous next week --> | ||
+ | <br><br> | ||
+ | <table border="0" width="100%" class="menu"> | ||
+ | <tr> | ||
+ | <td align="left">[[Team:UNIPV-Pavia/Calendar/June/settimana3| Previous week]]</td> | ||
+ | <td align="right">[[Team:UNIPV-Pavia/Calendar/July/settimana1| Next week]]</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | <!-- fine table previous next week --> | ||
+ | </td> | ||
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+ | {{UNIPV-Pavia/menu_mesi}} | ||
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Latest revision as of 08:00, 31 August 2010
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