Team:UNIPV-Pavia/Calendar/June

From 2010.igem.org

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<tr><td align="center" valign="top">June, 3th</td><td align="left">9° Meeting - Discussion of final and technical details about [[Team:UNIPV-Pavia/Project|The Project]]. Planning of Bio-Lab activity.</td></tr>
<tr><td align="center" valign="top">June, 3th</td><td align="left">9° Meeting - Discussion of final and technical details about [[Team:UNIPV-Pavia/Project|The Project]]. Planning of Bio-Lab activity.</td></tr>
<tr><td align="center" valign="top">[[#June, 7th|June, 7th]]</td><td align="left">1° Bio-Lab - Susanna, Nicolò and Federica.<br> <partinfo>BBa_E2050</partinfo>, <partinfo>BBa_K165018</partinfo>, <partinfo>BBa_K165037</partinfo>, <partinfo>BBa_J61001</partinfo>, <partinfo>BBa_K081008</partinfo> and <partinfo>BBa_K125500</partinfo> were resuspended from Spring 2010 DNA distribution. <partinfo>BBa_P1004</partinfo> was resuspended from Spring 2009 DNA distribution. All BioBricks were transformed in E. coli DH5alpha.<br>Liquid LB+Amp and LB agar plates +Amp were prepared according with "protocols".</td></tr>
<tr><td align="center" valign="top">[[#June, 7th|June, 7th]]</td><td align="left">1° Bio-Lab - Susanna, Nicolò and Federica.<br> <partinfo>BBa_E2050</partinfo>, <partinfo>BBa_K165018</partinfo>, <partinfo>BBa_K165037</partinfo>, <partinfo>BBa_J61001</partinfo>, <partinfo>BBa_K081008</partinfo> and <partinfo>BBa_K125500</partinfo> were resuspended from Spring 2010 DNA distribution. <partinfo>BBa_P1004</partinfo> was resuspended from Spring 2009 DNA distribution. All BioBricks were transformed in E. coli DH5alpha.<br>Liquid LB+Amp and LB agar plates +Amp were prepared according with "protocols".</td></tr>
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<tr><td align="center" valign="top">[[#June, 8th|June, 8th]]</td><td align="left">2° Bio-Lab - Susanna, Alessandro and Chiara.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 8th|June, 8th]]</td><td align="left">2° Bio-Lab - Susanna Z., Alessandro and Chiara.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 9th|June, 9th]]</td><td align="left">3° Bio-Lab - Susanna, Nicolò and Sara.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 9th|June, 9th]]</td><td align="left">3° Bio-Lab - Susanna Z., Nicolò and Sara.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 10th|June, 10th]]</td><td align="left">4° Bio-Lab - Susanna, Manuel Z. and Riccardo.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 10th|June, 10th]]</td><td align="left">4° Bio-Lab - Susanna Z., Manuel Z. and Riccardo.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 11th|June, 11th]]</td><td align="left">5° Bio-Lab - Susanna and Nicolò.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 11th|June, 11th]]</td><td align="left">5° Bio-Lab - Susanna Z. and Nicolò.<br></td></tr>
<tr><td align="center" valign="top">June, 12th</td><td align="left">iGEM EU workshop<br></td></tr>
<tr><td align="center" valign="top">June, 12th</td><td align="left">iGEM EU workshop<br></td></tr>
<tr><td align="center" valign="top">June, 13th</td><td align="left">iGEM EU workshop<br></td></tr>
<tr><td align="center" valign="top">June, 13th</td><td align="left">iGEM EU workshop<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 14th|June, 14th]]</td><td align="left">6° Bio-Lab - Lorenzo and Nicolò.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 15th|June, 15th]]</td><td align="left">7° Bio-Lab - Lorenzo and Manuel L., Sara and Alessandro.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 16th|June, 16th]]</td><td align="left">8° Bio-Lab - Susanna Z., Sara and Nicolò.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 17th|June, 17th]]</td><td align="left">9° Bio-Lab - Susanna Z., Nicolò and Riccardo.<br></td></tr>
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<tr><td align="center" valign="top">[[#June, 18th|June, 18th]]</td><td align="left">10° Bio-Lab - Susanna Z., Susanna S. and Federica.<br></td></tr>
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</table>
</table>
==June, 7th==
==June, 7th==
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==June, 11th==
==June, 11th==
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==June, 14th==
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==June, 15th==
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==June, 16th==
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==June, 17th==
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==June, 18th==

Revision as of 12:55, 17 June 2010
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JUNE


DATEACTIVITY
June, 3th9° Meeting - Discussion of final and technical details about The Project. Planning of Bio-Lab activity.
June, 7th1° Bio-Lab - Susanna, Nicolò and Federica.
<partinfo>BBa_E2050</partinfo>, <partinfo>BBa_K165018</partinfo>, <partinfo>BBa_K165037</partinfo>, <partinfo>BBa_J61001</partinfo>, <partinfo>BBa_K081008</partinfo> and <partinfo>BBa_K125500</partinfo> were resuspended from Spring 2010 DNA distribution. <partinfo>BBa_P1004</partinfo> was resuspended from Spring 2009 DNA distribution. All BioBricks were transformed in E. coli DH5alpha.
Liquid LB+Amp and LB agar plates +Amp were prepared according with "protocols".
June, 8th2° Bio-Lab - Susanna Z., Alessandro and Chiara.
June, 9th3° Bio-Lab - Susanna Z., Nicolò and Sara.
June, 10th4° Bio-Lab - Susanna Z., Manuel Z. and Riccardo.
June, 11th5° Bio-Lab - Susanna Z. and Nicolò.
June, 12thiGEM EU workshop
June, 13thiGEM EU workshop
June, 14th6° Bio-Lab - Lorenzo and Nicolò.
June, 15th7° Bio-Lab - Lorenzo and Manuel L., Sara and Alessandro.
June, 16th8° Bio-Lab - Susanna Z., Sara and Nicolò.
June, 17th9° Bio-Lab - Susanna Z., Nicolò and Riccardo.
June, 18th10° Bio-Lab - Susanna Z., Susanna S. and Federica.

June, 7th

These BioBrick were resuspended in 15ul ddH20:

  • BBa_E2050 (mOrange, 744bp) 2010 Kit Plate 2, well 13N in pSB2K3
  • BBa_K165018 (ADH1 terminator, 253 bp) 2010 Kit Plate 3, well 2M, J63009 (Amp)
  • BBa_K165037 (tef2 promoter, 403bp) 2010 Kit Plate 3, well 22O, pSB1AK3
  • BBa_P1004 (Chloramphenicol resistence cassette, 769 bp) 2009 Kit plate 1, well 7B, pSB1A1
  • BBa_J61001 (R6K origin, 406bp) 2010 Kit plate 1, well 240, pSB1A2
  • BBa_K081008 (RBS-luxI, 664bp) 2010 Kit plate 2, well 10L, pSB1A2
  • BBa_K125500 (GFP fusion brick, 718bp) 2010 Kit plate 3, well 2P, pSB1A2

1,5 ul of each culture was transformed in 100 ul of home-made competent E. coli DH5alpha.

Tranformants were all plated on LB agar plates added with Ampicillin, except for BBa_E2050 (Kanamycin).

500 ml LB+Amp was prepared and 21 LB agar plates + Amp were prepared (500 ml).


June, 8th

All plates grown overnight at 37°C show colonies! In particular, <partinfo>BBa_E2050</partinfo> (gorwn on LB+Kan), <partinfo>BBa_K125500</partinfo> (gorwn on LB+Amp), <partinfo>BBa_K081008</partinfo> (gorwn on LB+Amp) and <partinfo>BBa_K165018</partinfo> (gorwn on LB+Amp) showed big colonies, well separated on the plate. <partinfo>BBa_P1004</partinfo> (gorwn on LB+Amp) and <partinfo>BBa_J61001</partinfo> (gorwn on LB+Amp) showed many colonies, with big colonies surrounded by small colonies. <partinfo>BBa_K165037</partinfo> (gorwn on LB+Amp) showed only 11 big colonies.

A colony was peaked for each plate and inoculated in 1ml LB+antibiotic.


<partinfo>BBa_E2050</partinfo> 1ml LB+Kan
<partinfo>BBa_K165037</partinfo> 1ml LB+Amp
<partinfo>BBa_P1004</partinfo> 1ml LB+Amp
<partinfo>BBa_K125500</partinfo> 1ml LB+Amp
<partinfo>BBa_K081008</partinfo> 1ml LB+Amp
<partinfo>BBa_J61001</partinfo> 1ml LB+Amp
<partinfo>BBa_K165018</partinfo> 1ml LB+Amp
Didascalia2


<partinfo>BBa_K165037</partinfo> in pSB1AK3 plasmid was inocultaed both in 1ml LB+Amp and in 1ml LB+Kan for a phenotipic assay.

Cultures were grown for 8 hours at 37°C 220 rpm.

After this time, cultures were all grown and in saturation phase.

Glycerol stocks were prepared for each culture (250ul saturated culture + 750 ul glycerol 80%) and stored at -80°C.

The left culture was re-filled to 5ml with LB+antibiotic and grown overnight at 37°C 200 rpm to be miniprepped tomorrow.

June, 9th

Ligation of:

I0: <partinfo>BBa_K125500</partinfo> (E-S)+<partinfo>BBa_B0015</partinfo> (E-X) I1: <partinfo>BBa_E2050</partinfo> (E-S)+<partinfo>BBa_K165018</partinfo> (E-X) I2: <partinfo>BBa_P1004</partinfo> (E-S)+<partinfo>BBa_B0015</partinfo> (E-X) I3: <partinfo>BBa_K081008</partinfo> (E-S)+<partinfo>BBa_B0015</partinfo> (E-X)


<partinfo>BBa_B0015</partinfo> was accidentally not inoculated yesterday... :( Luckily, we had a DNA stock for this part stored at -20°C, so we used this stock for ligations.

Cultures grown overnight at 37°C 220 rpm were all saturated. BioBricks were extracted with MiniPrep kit. Surprisingly, after the first centrifugation, we observed that <partinfo>BBa_E2050</partinfo> pellet was orange!! This is curious, because this BioBrick expresses mOrange, but it lacks of promoters. Probably, this spurious transcription is due to the promoter of Kanamycin resistance, that is oriented in the same direction.

BBa_E2050 orange pellet


After MiniPrep, purified DNA was quantified with NanoDrop.

<partinfo>BBa_E2050</partinfo> 139,2 ng/ul
<partinfo>BBa_K165037</partinfo> 181,5 ng/ul
<partinfo>BBa_P1004</partinfo> 132,8 ng/ul
<partinfo>BBa_K125500</partinfo> 124,3 ng/ul
<partinfo>BBa_K081008</partinfo> 138,3 ng/ul
<partinfo>BBa_J61001</partinfo> 122,2 ng/ul
<partinfo>BBa_K165018</partinfo> 166,3 ng/ul

Digestion of:

Culture Kind Final reaction volume (ul) DNA (ul) H20 (ul) Enzyme 1 Enzyme 2 Buffer H
<partinfo>BBa_K125500</partinfo> Insert 16 4,5 1 EcoRI 1 SpeI 2,5
<partinfo>BBa_P1004</partinfo> Insert 16 4,5 1 EcoRI 1 SpeI 2,5
<partinfo>BBa_K125500</partinfo> 124,3 ng/ul
<partinfo>BBa_K081008</partinfo> 138,3 ng/ul
<partinfo>BBa_J61001</partinfo> 122,2 ng/ul
<partinfo>BBa_K165018</partinfo> 166,3 ng/ul

June, 10th

June, 11th

June, 14th

June, 15th

June, 16th

June, 17th

June, 18th


CALENDAR
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April
May
June
July
August
September
October
November

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