Team:UNIPV-Pavia/Calendar/June

From 2010.igem.org

(Difference between revisions)
(→June, 9th)
 
(81 intermediate revisions not shown)
Line 2: Line 2:
<table width="100%" border="0">
<table width="100%" border="0">
<tr>
<tr>
-
       <td colspan="2"> {{UNIPV-Pavia/header}} </td>   
+
       <td colspan="3"> {{UNIPV-Pavia/header}} </td>   
-
         <!-- <td colspan="2" align="center"><html><img src="http://img337.imageshack.us/img337/7634/logoigem.jpg"  width="100%" /></html></td> -->  
+
         <!-- <td colspan="3" align="center"><html><img src="http://img337.imageshack.us/img337/7634/logoigem.jpg"  width="100%" /></html></td> -->  
</tr>
</tr>
{{UNIPV-Pavia/Style}}
{{UNIPV-Pavia/Style}}
-
<tr><td align="left" valign="top" width="20%">{{UNIPV-Pavia/menu}}</td>
+
<tr><td align="left" valign="top" width="15%">{{UNIPV-Pavia/menu}}</td>
<td valign="top">
<td valign="top">
-
<table border="0" align="center" width="100%"><tr><td align="justify" valign="top">
+
<table border="0" align="center" width="100%" class="trasparenza"><tr><td align="justify" valign="top" style="padding:20px">
-
<html><p align="center"><font size="4"><b>JUNE</b></font></p></html><hr><br>
+
-
<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><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, 8th|June, 8th]]</td><td align="left">2° Bio-Lab - Susanna, Alessandro and Chiara.<br></td></tr>
+
-
<tr><td align="center" valign="top">[[#June, 9th|June, 9th]]</td><td align="left">3° Bio-Lab - Susanna, Nicolò and Sara.<br></td></tr>
+
-
<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>
+
-
<tr><td align="center" valign="top">[[#June, 11th|June, 11th]]</td><td align="left">5° Bio-Lab - Susanna 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, 13th</td><td align="left">iGEM EU workshop<br></td></tr>
+
-
</table>
+
-
==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.
+
-
 
+
-
<center>
+
<table class="menu" border="0" width="100%">
-
<table width='100%'>
+
<tr>
<tr>
-
<td>
+
<td align="center">
-
{|
+
[[Team:UNIPV-Pavia/Calendar/June/settimana2|Week 2]]
-
|  <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 
+
-
|}
+
</td>
</td>
-
<td>
+
<td align="center">
-
<table>
+
[[Team:UNIPV-Pavia/Calendar/June/settimana3|Week 3]]
-
<tr>
+
-
<td>
+
-
[[Image:UNIPV_attivita_lab1.jpg|thumb|300px|left|Didascalia1]]
+
</td>
</td>
-
</tr>
+
<td align="center">
-
<tr>
+
[[Team:UNIPV-Pavia/Calendar/June/settimana4|Week 4]]
-
<td>
+
-
[[Image:UNIPV_attivita_lab2.jpg|thumb|300px|left|Didascalia2]]
+
</td>
</td>
</tr>
</tr>
 +
</table>
 +
 +
 +
<html><p align="center"><font size="4"><b>JUNE</b></font></p></html><hr><br>
 +
Finally Bio-Lab activity began!<br><br><font size="4"><b><u> Select the day you are interested in to find out all the details!</u></b></font><br><br>
 +
<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><td align="center" valign="top"><html><a name="June_3rd"></a></html>June, 3rd</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><th align="center" width="12%" colspan="2"><b>JUNE: WEEK 2</b></th></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana2 #June, 7th|June, 7th]]</td><td align="left">1° Bio-Lab - <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+Amp agar plates were prepared according with "protocols".</td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana2 #June, 8th|June, 8th]]</td><td align="left">2° Bio-Lab - Inoculum from plates in liquid Lb and glycerol stocks preparation. Falcon were re-filled for tomorrow mini-prep.<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana2 #June, 9th|June, 9th]]</td><td align="left">3° Bio-Lab - MiniPrep of <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>. Digestion, gel run/cut and 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)<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana2 #June, 10th|June, 10th]]</td><td align="left">4° Bio-Lab - Transformation of ligations I0, I1, I2, I3 in home made ''E. coli'' DH5-alpha competent cells. Transformants were plated on LB+Amp agar plates.<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana2 #June, 11th|June, 11th]]</td><td align="left">5° Bio-Lab - Inoculum from plates in liquid LB and glycerol stocks preparation for I0, I1, I2 and I3 in duplicate (2 colonies picked from each plate).<br></td></tr>
 +
<tr><td align="center" valign="top"><html><a name="June_12th"></a></html>June, 12th</td><td align="left">iGEM EU workshop.<br></td></tr>
 +
<tr><td align="center" valign="top"><html><a name="June_13th"></a></html>June, 13th</td><td align="left">iGEM EU workshop.<br></td></tr>
 +
<tr><th align="center" width="12%" colspan="2"><b>JUNE: WEEK 3</b></th></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana3 #June, 14th|June, 14th]]</td><td align="left">6° Bio-Lab -  Inoculum from glycerol stock of I0-1, I0-2, I1-1, i1-2, I2-1, I3-1 and I3-2 for screening.</td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana3 #June, 15th|June, 15th]]</td><td align="left">7° Bio-Lab - MiniPrep, gel run/cut and ligation of
 +
*I4: <partinfo>BBa_K165037</partinfo> (S-P) + I1 (X-P)
 +
*I5: I2 (E-S) + <partinfo>BBa_J61001</partinfo> (E-X)
 +
*I6: I3 (E-S) + <partinfo>BBa_T9002</partinfo> (E-X)<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana3 #June, 16th|June, 16th]]</td><td align="left">8° Bio-Lab - Tranformation of I4 (DH5-alpha), I5 (TOP10) and I6 (DH5-alpha).<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana3 #June, 17th|June, 17th]]</td><td align="left">9° Bio-Lab - Colonies were picked from each plate, I4-1, I4-2, I5-1, I5-2, I6-1, I6-2 and I6-3 were grown in 1ml liquid culture and glycerol stocked (not I5-2, because no growth was observed in LB+Cm and not for I6-1, not grown). Cultures refilled with 5ml LB+antibiotic for tomorrow MiniPrep and screening.<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana3 #June, 18th|June, 18th]]</td><td align="left">10° Bio-Lab - MiniPrep for I4-1, I4-2, I5-1, I6-2 and I6-3. Screening gel (all the clones are ok!) :)<br></td></tr>
 +
<tr><th align="center" width="12%" colspan="2"><b>JUNE: WEEK 4</b></th></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana4 #June, 21st|June, 21st]]</td><td align="left">11° Bio-Lab - Week activities planning and freezer sorting.<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana4 #June, 22nd|June, 22nd]]</td><td align="left">12° Bio-Lab - Inoculations of I6, BBa_J23118, BBa_J23110, BBa_J23114, BBa_J23116.<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana4 #June, 23rd|June, 23rd]]</td><td align="left">13° Bio-Lab - 10° Meeting - Updating about Bio-Lab activity.<br>MiniPrep of ON cultures I6, BBa_J23118, BBa_J23110, BBa_J23114, BBa_J23116, with NanoDrop quantification. Digestions, gel run and extraction of the above. We received BBa_K208001, the Silver-fusion compatible BioBrick part that codes for phasin. Streaked them on two LB+Kan agar plates (named PhaP-1 and PhaP-2)</td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana4 #June, 24th|June, 24th]]</td><td align="left">14° Bio-Lab - Colonies on both PhaP-1 and PhaP-2. Inoculation of one colony per plate, with subsequent growth for 6hrs. Glycerol stocks and refill for tomorrow MiniPrep. Yesterday's ligations heated to inactivate ligase and transformed in TOP10 homemade competent cells.<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/June/settimana4 #June, 25th|June, 25th]]</td><td align="left">15° Bio-Lab - Plates all show colonies! Picked two colonies from each plate and incubated in 1ml LB+Amp. The cultures were grown for six hours then glycerol stocks were made. PhaP-1 and 2 MiniPrep and quantification by NanoDrop. DNA quantification was poor, so we decided to perform digestion screening and to repeat Miniprep next week for sequencing. Then they were digested and gel screened, both resulting positive. <br></td></tr>
 +
<tr><th align="center" width="12%" colspan="2"><b>JULY: WEEK 1</b></th></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana1 #June, 28th|June, 28th]]</td><td align="left">16° Bio-Lab - Inoculum from glycerol stock for I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 in 5ml LB+Amp. Cultures grown overnight. Inoculum of PhaP-1 and PhaP-2 in 5ml LB+Kan and ON growth. Received strains and plasmid from CGSC (Yale University). Details in the week view.<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana1 #June, 29th|June, 29th]]</td><td align="left">17° Bio-Lab - From I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 5ul were aliquoted and diluted in 500ul LB+Amp for a preliminary TECAN test. Then cultures were MiniPrepped. Almost all cultures were digested. PhaP-1 and PhaP-2 were prepared or sequencing. All strains received from Yale Univesity were on blotting paper disks, that were placed on an LB agar plates, resuspended and then streaked. After plate streaking, disks were transferred in falcon tubes containing liquid LB.<br></td></tr>
 +
<tr><td align="center" valign="top">[[Team:UNIPV-Pavia/Calendar/July/settimana1 #June, 30th|June, 30th]]</td><td align="left">18° Bio-Lab - Inoculum of cultures for a TECAN test.<br></td></tr>
</table>
</table>
</td>
</td>
</tr>
</tr>
</table>
</table>
-
</center>
 
-
 
-
 
-
<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.
 
-
 
-
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 
 
-
|}
 
-
 
-
==June, 10th==
 
-
 
-
==June, 11th==
 
-
 
-
 
-
 
-
</td>
 
-
<td width="20%" valign="top">
 
-
 
 +
<td width="15%" align="right" valign="top">
{{UNIPV-Pavia/menu_mesi}}
{{UNIPV-Pavia/menu_mesi}}

Latest revision as of 13:55, 20 October 2010
Home
The Project
Materials & Methods
Parts
The Team
Notebook
Biological Safety
Pavia
Gallery
Sponsors

Week 2

Week 3

Week 4


JUNE


Finally Bio-Lab activity began!

Select the day you are interested in to find out all the details!

DATEACTIVITY
June, 3rd9° Meeting - Discussion of final and technical details about The Project. Planning of Bio-Lab activity.
JUNE: WEEK 2
June, 7th1° Bio-Lab - <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+Amp agar plates were prepared according with "protocols".
June, 8th2° Bio-Lab - Inoculum from plates in liquid Lb and glycerol stocks preparation. Falcon were re-filled for tomorrow mini-prep.
June, 9th3° Bio-Lab - MiniPrep of <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>. Digestion, gel run/cut and 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)
June, 10th4° Bio-Lab - Transformation of ligations I0, I1, I2, I3 in home made E. coli DH5-alpha competent cells. Transformants were plated on LB+Amp agar plates.
June, 11th5° Bio-Lab - Inoculum from plates in liquid LB and glycerol stocks preparation for I0, I1, I2 and I3 in duplicate (2 colonies picked from each plate).
June, 12thiGEM EU workshop.
June, 13thiGEM EU workshop.
JUNE: WEEK 3
June, 14th6° Bio-Lab - Inoculum from glycerol stock of I0-1, I0-2, I1-1, i1-2, I2-1, I3-1 and I3-2 for screening.
June, 15th7° Bio-Lab - MiniPrep, gel run/cut and ligation of
  • I4: <partinfo>BBa_K165037</partinfo> (S-P) + I1 (X-P)
  • I5: I2 (E-S) + <partinfo>BBa_J61001</partinfo> (E-X)
  • I6: I3 (E-S) + <partinfo>BBa_T9002</partinfo> (E-X)
June, 16th8° Bio-Lab - Tranformation of I4 (DH5-alpha), I5 (TOP10) and I6 (DH5-alpha).
June, 17th9° Bio-Lab - Colonies were picked from each plate, I4-1, I4-2, I5-1, I5-2, I6-1, I6-2 and I6-3 were grown in 1ml liquid culture and glycerol stocked (not I5-2, because no growth was observed in LB+Cm and not for I6-1, not grown). Cultures refilled with 5ml LB+antibiotic for tomorrow MiniPrep and screening.
June, 18th10° Bio-Lab - MiniPrep for I4-1, I4-2, I5-1, I6-2 and I6-3. Screening gel (all the clones are ok!) :)
JUNE: WEEK 4
June, 21st11° Bio-Lab - Week activities planning and freezer sorting.
June, 22nd12° Bio-Lab - Inoculations of I6, BBa_J23118, BBa_J23110, BBa_J23114, BBa_J23116.
June, 23rd13° Bio-Lab - 10° Meeting - Updating about Bio-Lab activity.
MiniPrep of ON cultures I6, BBa_J23118, BBa_J23110, BBa_J23114, BBa_J23116, with NanoDrop quantification. Digestions, gel run and extraction of the above. We received BBa_K208001, the Silver-fusion compatible BioBrick part that codes for phasin. Streaked them on two LB+Kan agar plates (named PhaP-1 and PhaP-2)
June, 24th14° Bio-Lab - Colonies on both PhaP-1 and PhaP-2. Inoculation of one colony per plate, with subsequent growth for 6hrs. Glycerol stocks and refill for tomorrow MiniPrep. Yesterday's ligations heated to inactivate ligase and transformed in TOP10 homemade competent cells.
June, 25th15° Bio-Lab - Plates all show colonies! Picked two colonies from each plate and incubated in 1ml LB+Amp. The cultures were grown for six hours then glycerol stocks were made. PhaP-1 and 2 MiniPrep and quantification by NanoDrop. DNA quantification was poor, so we decided to perform digestion screening and to repeat Miniprep next week for sequencing. Then they were digested and gel screened, both resulting positive.
JULY: WEEK 1
June, 28th16° Bio-Lab - Inoculum from glycerol stock for I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 in 5ml LB+Amp. Cultures grown overnight. Inoculum of PhaP-1 and PhaP-2 in 5ml LB+Kan and ON growth. Received strains and plasmid from CGSC (Yale University). Details in the week view.
June, 29th17° Bio-Lab - From I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 5ul were aliquoted and diluted in 500ul LB+Amp for a preliminary TECAN test. Then cultures were MiniPrepped. Almost all cultures were digested. PhaP-1 and PhaP-2 were prepared or sequencing. All strains received from Yale Univesity were on blotting paper disks, that were placed on an LB agar plates, resuspended and then streaked. After plate streaking, disks were transferred in falcon tubes containing liquid LB.
June, 30th18° Bio-Lab - Inoculum of cultures for a TECAN test.

CALENDAR
March
April
May
June
July
August
September
October
November

Retrieved from "http://2010.igem.org/Team:UNIPV-Pavia/Calendar/June"