Team:UNIPV-Pavia/Calendar/August
From 2010.igem.org
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</table> | </table> | ||
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- | <html><p align="center"><font size="4"><b>AUGUST</b></font></p></html><hr | + | <html><p align="center"><font size="4"><b>AUGUST</b></font></p></html><hr><br> |
- | Bio-Lab activity proceeds!<br> | + | Bio-Lab activity proceeds!<br><br> |
- | <font size = "4"><b><u>Select the day you are interested to find out the details!</u></b></font><br><br> | + | <font size = "4"><b><u>Select the day you are interested in to find out the details!</u></b></font><br><br> |
<table class=cont border="0" width="30%" align="center" valign="top"> | <table class=cont border="0" width="30%" align="center" valign="top"> | ||
<tr><th align="center" width="12%"><b>DATE</b></th><th align="center"><b>ACTIVITY</b></th></tr> | <tr><th align="center" width="12%"><b>DATE</b></th><th align="center"><b>ACTIVITY</b></th></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #August, 1st|August, 1st]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana5 #August, 1st|August, 1st]]</td><td align="left"> | ||
- | + | 45° Bio-Lab - I20 and I21 plates showed colonies. Some were picked, inoculated and grown in proper medium. | |
</td></tr> | </td></tr> | ||
<tr><th align="center" width="12%" colspan="2"><b>AUGUST: WEEK 1</b></th></tr> | <tr><th align="center" width="12%" colspan="2"><b>AUGUST: WEEK 1</b></th></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 2nd|August, 2nd]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 2nd|August, 2nd]]</td><td align="left"> | ||
- | + | 46° Bio-Lab - I20-1/2/3 and I21-1/2/3 ligations sent sequencing. | |
<partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081009</partinfo> resuspended from iGEM 2010 Distribution Kit. | <partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081009</partinfo> resuspended from iGEM 2010 Distribution Kit. | ||
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</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 3rd|August, 3rd]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 3rd|August, 3rd]]</td><td align="left"> | ||
- | + | 47° Bio-Lab - | |
<partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081009</partinfo> single colony pick from plates. | <partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081009</partinfo> single colony pick from plates. | ||
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</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 4th|August, 4th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 4th|August, 4th]]</td><td align="left"> | ||
- | + | 48° Bio-Lab - Glycerol stocks for MC1061 and MG1655 strains transformed with pAH123 or <partinfo>BBa_J72008</partinfo> helper plasmids. Miniprep of MC1061 and MG1655 to check again the presence of helper plasmids. Digestion with SpeI and gel run. We got positive samples but a bad run. So we picked again from plates, this time single colonies. | |
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 5th|August, 5th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 5th|August, 5th]]</td><td align="left"> | ||
- | + | 49° Bio-Lab - Screening for MG1655 and MC1061 carrying <partinfo>BBa_J72008</partinfo> or pAH123 helpers: miniprep of new picked colonies MC1061 and MG1655, digestion with SpeI and gel run. | |
- | + | ||
+ | New ligation cycle for phasins (I20-new, I21-new). | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 6th|August, 6th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 6th|August, 6th]]</td><td align="left"> | ||
- | + | 50° Bio-Lab - Transformation of I20-new and I21-new ligations into ''E. coli Dh5-alpha''. | |
+ | |||
+ | Competentization of MG008/123 and MC008/123 strains selected previous day. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 7th|August, 7th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana1 #August, 7th|August, 7th]]</td><td align="left"> | ||
- | + | 51° Bio-Lab - Plates of transformed cells were stored at +4°C. A Tecan test was performed with usual protocol. | |
- | + | ||
- | + | ||
- | 51° Bio-Lab - | + | |
</td></tr> | </td></tr> | ||
<tr><th align="center" width="12%" colspan="2"><b>AUGUST: WEEK 2</b></th></tr> | <tr><th align="center" width="12%" colspan="2"><b>AUGUST: WEEK 2</b></th></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 9th|August, 9th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 9th|August, 9th]]</td><td align="left"> | ||
- | 52° Bio-Lab - | + | 52° Bio-Lab - Phasins plates showed very few colonies (17 for I20-new and 12 for I21-new): they were all picked and let grow in LB+Amp 100ug/ml. In the evening we made glycerol stocks and re-filled the tubes for the screening of the following day. Not to loose time we performed again PCR-amplification/modification of <partinfo>BBa_K208001</partinfo>. |
+ | |||
+ | We checked LB+Cm 6 ug/ml agar plates with MC1061, MG1655, MC008, MG008, MC123, MG123 (none of them should grow). | ||
+ | |||
+ | Check for transformation efficiency of MC123/008 and MG123/008. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 10th|August, 10th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 10th|August, 10th]]</td><td align="left"> | ||
- | 53° Bio-Lab - | + | 53° Bio-Lab - Glycerol stock was prepared for: linker (<partinfo>BBa_K105012</partinfo>), I22-1, I22-2, I22-3, I23-1, I23-2, I23-3, I8-5 D, I8-5 E and I8-5 F. |
+ | |||
+ | I8-5 D, I8-5 E, I8-5 F, I74C5, I84C5, I104C5, I124C5, <partinfo>BBa_B0032</partinfo>, A2 were diluited for Tecan Test. | ||
+ | |||
+ | MiniPrep and digestion was performed for following cultures: I20-1, I20-2, I20-3, I20-4, I20-5, I20-6, I20-7, I20-8, I20-9, I20-10, I20-11, I20-12, I21-1, I21-2, I21-3, I21-4, I21-5, I21-6, I21-7, I21-8, I21-9, I21-10, I21-11, I21-12, I21-13, I21-14, I21-15, I21-16, I21-17, I22-1, I22-2, I22-3, I23-1, I23-2, I23-3, I15-1, Ent4C5, <partinfo>BBa_B0034</partinfo>, linker (<partinfo>BBa_K105012</partinfo>) and plux (<partinfo>BBa_R0062</partinfo>). | ||
+ | Samples were gel ran/cut. After purification, digested DNA was quantified. | ||
+ | |||
+ | Ligation of: I15_4C5, I24, I26, I20_M, I21_M, I20_A, I21_A, I20_C, I21_C. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 11th|August, 11th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 11th|August, 11th]]</td><td align="left"> | ||
- | 54° Bio-Lab - | + | 54° Bio-Lab - Trasformation of ligations of yestrday. |
+ | |||
+ | The following plates were checked: F2620-4C5 in MC123, MC008, MG123, MG008;RING in MC123, MC008, MG123, MG008; NOTHING in MC123, MC008, MG123, MG008 | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 12th|August, 12th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 12th|August, 12th]]</td><td align="left"> | ||
- | 55° Bio-Lab - | + | 55° Bio-Lab - A massive screening for phasins was performed on I20_M-1, I21_M-2, I20_M-3, I20_M-4, I21_M-1, I21_M-2, I21_M-3, I21_M-4, I20_A-1, I20_A-2, I20_A-3, I20_A-4, I20_A-5, I21_A-6, I20_A-7, I21_A-1, I21_A-2, I21_A-3, I21_A-4, I21_A-5,I21_A-6, I21_A-7, I20_C-1, I20_C-2, I20_C-3, I20_C-4, I21_C-1, I21_C-2, I21_C-3, I21_C-4, I21_C-5, I26-1, I26-2, I26-3, I26-4 and I26-5. |
+ | |||
+ | For the cultures that Screening showed positive (colonies bearing the insert) an inoculum was performed in 6ml LB+Amp. | ||
+ | |||
+ | Glycerol stock was prepared for: I15_4C5-1, I15_4C5-2, I15_4C5-3, I24-1, I24-2 and I24-3. | ||
</td></tr> | </td></tr> | ||
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 13th|August, 13th]]</td><td align="left"> | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana2 #August, 13th|August, 13th]]</td><td align="left"> | ||
- | 56° Bio-Lab - | + | 56° Bio-Lab - Intein and CREAM were stored +4°C. |
+ | |||
+ | Efficiency of our home-made competent cells MC123/008 and MG123/008 wwas calculated. | ||
+ | |||
+ | Glycerol stock was prepared for: I20_M-1, I21_M-3, I20_A-1, I20_A-2, I20_A-3, I20_A-4, I20_A-6, I21_A-2, I21_A-3, I21_A-4, I21_A-5, I21_A-6, I21_A-7 and I26-2. | ||
+ | |||
+ | MiniPrep and digestion was performed for the following cultures: I15_4C5-1, I15_4C5-2, I15_4C5-3, I24-1, I24-2, I24-3, I20_M-1, I21_M-3, I20_A-1, I20_A-2, I20_A-3, I20_A-4, I20_A-6, I21_A-2, I21_A-3, I21_A-4, I21_A-5, I21_A-6, I21_A-7 and I26-2. | ||
+ | Cultures were digested and gel run. | ||
</td></tr> | </td></tr> | ||
- | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/ | + | <tr><th align="center" width="12%" colspan="2"><b>AUGUST: WEEK 3</b></th></tr> |
- | 57° Bio-Lab - | + | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana3 #August, 16th|August, 16th]]</td><td align="left"> |
+ | 57° Bio-Lab - Resuspension from iGEM 2010 Distribution Kit of <partinfo>BBa_J13002</partinfo> and <partinfo>BBa_I13522</partinfo>. | ||
+ | |||
+ | Transformation (1 ul) of <partinfo>BBa_J13002</partinfo>, <partinfo>BBa_I13522</partinfo>, INTEIN, CREAM (Mr.Gene) and 15_4C5. | ||
+ | |||
+ | Transfomants were plated on LB+Amp 100 ug/ml agar plates or on LB+Amp100+Cm12,5 agar plates and were let grow. | ||
</td></tr> | </td></tr> | ||
- | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/ | + | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana3 #August, 17th|August, 17th]]</td><td align="left"> |
- | 58° Bio-Lab - | + | 58° Bio-Lab - Pick of <partinfo>BBa_J13002</partinfo>, <partinfo>BBa_I13522</partinfo>, INTEIN, CREAM, co-trasformed <partinfo>BBa_T9002</partinfo>. |
+ | |||
+ | Inoculum of I5, I26, <partinfo>BBa_K105012</partinfo>, <partinfo>BBa_B0034</partinfo>, I20M-1, I20A-1, I21M-3, I21-4, I21A-7 and I0. | ||
+ | |||
+ | I7-A, I8-5 D, I9-1, I10-B, I12-2, A2, BBa_B0032, I144C5-T9002, I164C5-T9002, I174C5-T9002, I184C5-T9002, I194C5-T9002 and ENTERO4C5-T9002 were diluted for Tecan Test. | ||
</td></tr> | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana3 #August, 18th|August, 18th]]</td><td align="left"> | ||
+ | 59° Bio-Lab - I7-A, I8-5 D, I9-1, I10-B, I12-2, A2, BBa_B0032, I144C5-T9002, I154C5-T9002, I164C5-T9002, I174C5-T9002, I184C5-T9002, I194C5-T9002 and ENTERO4C5-T9002 were diluted for Tecan Test. | ||
+ | The following cultures were miniprepped and after digested: I26, BBa_K105012, BBa_B0034, BBa_F2620, I20M-1, I20A-1, I21M-3, I21-4, I0, INTEIN and BBa_J13002. | ||
+ | Two medium gel were prepared and samples were loaded and gel run/cut. After gel extraction they were stored at -20°. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana3 #August, 19th|August, 19th]]</td><td align="left"> | ||
+ | 60° Bio-Lab - After gel exctraction were performed the following ligations: I31, I32, I33, I34, I35, I36, I37, I38, I39, I40 and I41. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana3 #August, 20th|August, 20th]]</td><td align="left"> | ||
+ | 61° Bio-Lab - Ligations of previous day were transformed and than plated on agar plates and incubated. | ||
+ | |||
+ | A Tecan test was performed with the usual protocol. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana3 #August, 21st|August, 21st]]</td><td align="left"> | ||
+ | 62° Bio-Lab - Plates with ligations I31..I41 were stored at +4°C. | ||
+ | </td></tr> | ||
+ | <tr><th align="center" width="12%" colspan="2"><b>AUGUST: WEEK 4</b></th></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana4 #August, 23rd|August, 23rd]]</td><td align="left"> | ||
+ | 63° Bio-Lab - Colony PCR as screening for ligations I31..I41. For each plate three colonies were picked and PCR amplification was performed. | ||
+ | |||
+ | MyCrim-9, BBa_K173001, BBa_J23101, pSB4C5 were digested, after that gel exctraction was performed. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana4 #August, 24th|August, 24th]]</td><td align="left"> | ||
+ | 64° Bio-Lab - The following cultures :I32 (I32-4/5/6/7), I36 (I36-4/5/6/7) and I39 (I39/4/5/6/7) were screened. | ||
+ | |||
+ | Inoculum of <partinfo>BBa_J23105</partinfo>, <partinfo>BBa_J23106</partinfo>, <partinfo>BBa_J23114</partinfo> and <partinfo>BBa_J23116</partinfo>. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana4 #August, 25th|August, 25th]]</td><td align="left"> | ||
+ | 65° Bio-Lab -Miniprep was performed for I40-1/4/5/6/7 to make the screening of ligations. Samples were digested and than gel run to check the length of ligations. | ||
+ | |||
+ | MiniPrep was performed for <partinfo>BBa_J23100</partinfo>, <partinfo>BBa_J23105</partinfo>, <partinfo>BBa_J23106</partinfo>, <partinfo>BBa_J23110</partinfo>, <partinfo>BBa_J23114</partinfo>, <partinfo>BBa_J23116</partinfo> and <partinfo>BBa_J23118</partinfo> | ||
+ | |||
+ | Ligation of <partinfo>BBa_J23100</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 and <partinfo>BBa_J23118</partinfo>_4C5. | ||
+ | |||
+ | Preparation of samples for BioPlastic screening. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana4 #August, 26th|August, 26th]]</td><td align="left"> | ||
+ | 66° Bio-Lab - Transformation of ligations: <partinfo>BBa_J23100</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 and <partinfo>BBa_J23118</partinfo>_4C5. | ||
+ | |||
+ | Inoculum of self inducible promoters for TECAN test. | ||
+ | |||
+ | Inoculum of <partinfo>BBa_J13002</partinfo>, I33, I34, I39 and I41. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana4 #August, 27th|August, 27th]]</td><td align="left"> | ||
+ | 67° Bio-Lab - Inoculated strains (<partinfo>BBa_J13002</partinfo>, I33, I34, I39 and I41) were MiniPrepped and quantified.After digestion they were gel run and cut.Gel extraction was quantified nd ligation of I-47, I-48 and I-49 were performed. | ||
+ | Colony PCR was performed to screen if the length of insert was correct on <partinfo>BBa_J23100</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 and <partinfo>BBa_J23118</partinfo>_4C5. | ||
+ | |||
+ | Tecan Test was performed on prepared samples, after the usual protocol. | ||
+ | |||
+ | Transformation of I29 in E. coli TOP10. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/August/settimana4 #August, 28th|August, 28th]]</td><td align="left"> | ||
+ | 68° Bio-Lab - Ligations I47, I48, I49 and I29 plate were stored at +4°C. | ||
+ | </td></tr> | ||
+ | <tr><th align="center" width="12%" colspan="2"><b>SEPTEMBER: WEEK 1</b></th></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana1 #August, 30th|August, 30th]]</td><td align="left"> | ||
+ | 69° Bio-Lab - Colony PCR was performed on <partinfo>BBa_J23116</partinfo>_4C5 (3 colonies) and <partinfo>BBa_J23118</partinfo>_4C5 (3 colonies). | ||
+ | |||
+ | Transformation of ligations I47, I48, I49 into E. coli DH5-alpha. | ||
+ | </td></tr> | ||
+ | <tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/September/settimana1 #August, 31st|August, 31st]]</td><td align="left"> | ||
+ | 70° Bio-Lab - Glycerol stocks were prepared for I29_1, I29_2 and I29_3. | ||
+ | |||
+ | I29_1, I29_2 and I29_3 was MiniPrepped to purify DNA, and than gel was run to check the length of ligations. | ||
+ | |||
+ | Sudan Black staining protocol for BioPlastic was performed. | ||
+ | |||
+ | PCR from colony for I47, I48, I49 ligations was performed. | ||
+ | </td></tr> | ||
</table> | </table> |
Latest revision as of 11:01, 20 October 2010
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