Team:Paris Bettencourt/Notebook/Interlab Study

From 2014.igem.org

(Difference between revisions)
Line 1: Line 1:
-
{{Team:Paris_Bettencourt/Wiki}}
+
{{CSS/Main}}
-
{{Team:Paris_Bettencourt/Menu}}
+
<html>
<html>
-
<style>
+
<style>
-
 
+
-
#topheader {
+
-
        margin-left : 20%;
+
-
        width : 60%;
+
-
        height : 80px;
+
-
        margin-top : 40px;
+
-
}
+
#PBlogo {
#PBlogo {
float : left;
float : left;
Line 16: Line 8:
margin-left : 23%;
margin-left : 23%;
}
}
 +
 +
#iGEMlogo {
 +
width : 122px;
 +
float : right;
 +
margin-right : 23%;
 +
}
 +
 +
                h1 {
 +
                        font-size : 0px;
 +
                }
h2 {
h2 {
font-family : Times;
font-family : Times;
font-size : 30px;
font-size : 30px;
 +
text-decoration : underline;
text-align : center;
text-align : center;
}
}
Line 38: Line 41:
}
}
-
+
#cadre {
-
a {
+
border:5px double rgb(0,0,0);
 +
align="center";
 +
height ="45px";
 +
color: rgb(0,0,0);
 +
}
 +
 +
.a {
color : rgb(0,0,0);
color : rgb(0,0,0);
}
}
-
 
+
 +
#menu {
 +
text-align : center;
 +
height : 45px;
 +
width : 65%;
 +
margin-left : 17.5%;
 +
margin-right : 17.5%;
 +
}
 +
 +
#shadow {
#shadow {
-
margin-left :-50%;
+
margin-left : 25px;
float : left;
float : left;
border-radius : 5px;
border-radius : 5px;
Line 125: Line 143:
<div id="topheader">
<div id="topheader">
<img id="PBlogo" src="https://static.igem.org/mediawiki/2013/3/3a/PB_logoParis.gif"/>
<img id="PBlogo" src="https://static.igem.org/mediawiki/2013/3/3a/PB_logoParis.gif"/>
 +
<a href="https://2014.igem.org/Main_Page"> <img id="iGEMlogo" src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a>
 +
<h1></h1>
<h2>Paris Bettencourt 2014</h2>
<h2>Paris Bettencourt 2014</h2>
</div>
</div>
-
 
+
<table id=menu>
 +
<tr>
 +
<td id = "cadre"><a href="https://2014.igem.org/Team:Paris_Bettencourt">Home </a> </td>
 +
 +
<td id = "cadre"><a href="https://2014.igem.org/Team:Paris_Bettencourt/Team"> Team </a> </td>
 +
 +
<td id = "cadre"><a href="https://igem.org/Team.cgi?year=2014&team_name=Paris_Bettencourt"> Official Team Profile </a></td>
 +
 +
<td id = "cadre"><a href="https://2014.igem.org/Team:Paris_Bettencourt/Project"> Project</a></td>
 +
 +
<td id = "cadre"><a href="https://2014.igem.org/Team:Paris_Bettencourt/Parts"> Parts</a></td>
 +
 +
<td id = "cadre"><a href="https://2014.igem.org/Team:Paris_Bettencourt/Modeling"> Modeling</a></td>
 +
 +
<td id = "cadre"><a href="https://2014.igem.org/Team:Paris_Bettencourt/Notebook"> Notebook</a></td>
 +
 +
<td id = "cadre"><a href="https://2014.igem.org/Team:Paris_Bettencourt/Safety"> Safety </a></td>
 +
 +
<td id = "cadre"><a href="https://2014.igem.org/Team:Paris_Bettencourt/Attributions"> Attributions </a></td>
 +
</tr>
 +
</table>
</head>
</head>
Line 219: Line 259:
<h4>July </h4>
<h4>July </h4>
<h5 id="date">July 4th</h5>
<h5 id="date">July 4th</h5>
-
<p id="text">We will participate in the<a href=”https://2014.igem.org/Tracks/Measurement/Interlab_study” >  iGEM interlab study</a></p>
+
<p id="text">We will participate in the<a href="https://2014.igem.org/Tracks/Measurement/Interlab_study" >  iGEM interlab study</a></p>
<p><b><u>Device 1</b></u></p>
<p><b><u>Device 1</b></u></p>
Line 289: Line 329:
</ul></p>
</ul></p>
-
<p> For the <u>successful transformations</u> I followed the <a href=https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">Glycerol stock </a> protocol </p>
+
<p> For the <u>successful transformations</u> I followed the <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">Glycerol stock </a> protocol </p>
<p>For the <u> unsuccessful transformation </u> I transformed NEB turbo again with the rest of Biobrick BBa_K823005 following again <a href=kit>iGEM Distribution Kit instructions </a> and <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1">Heat Shock transformation of <i>E.coli</i></a></p>
<p>For the <u> unsuccessful transformation </u> I transformed NEB turbo again with the rest of Biobrick BBa_K823005 following again <a href=kit>iGEM Distribution Kit instructions </a> and <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1">Heat Shock transformation of <i>E.coli</i></a></p>
Line 296: Line 336:
<h5 id="date">July 9th</h5>
<h5 id="date">July 9th</h5>
-
<p> I finished the <a href=https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">Glycerol stock </a> protocol and labelled G.23 for BBa_K823012 and G.24 for BBa_E0240</p>
+
<p> I finished the <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">Glycerol stock </a> protocol and labelled G.23 for BBa_K823012 and G.24 for BBa_E0240</p>
-
<p><b>BBa_K823005 </b> successfully transformed! I followed the <a href=https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">Glycerol stock </a> protocol </p>
+
<p><b>BBa_K823005 </b> successfully transformed! I followed the <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">Glycerol stock </a> protocol </p>
<p><u>Minipreps</u>
<p><u>Minipreps</u>
</br>
</br>
-
Following the standard  <a href=https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot4">protocol fot the miniprep  </a></br>
+
Following the standard  <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot4">protocol fot the miniprep  </a></br>
I measured DNA content with the nanodrop. </br>
I measured DNA content with the nanodrop. </br>
<u>Digestion analysis</u>: (Clovis's recipe)</br>
<u>Digestion analysis</u>: (Clovis's recipe)</br>
Line 317: Line 357:
<h5 id="date">July 10th</h5>
<h5 id="date">July 10th</h5>
-
<p>Finished the <a href=https://2014.igem.org/Team:Paris_Betmisslabelledtencourt/Protocols#prot8">Glycerol stock </a> protocol and labelled it G.25</p>
+
<p>Finished the <a href="https://2014.igem.org/Team:Paris_Betmisslabelledtencourt/Protocols#prot8">Glycerol stock </a> protocol and labelled it G.25</p>
<p><u> Minipreps</u>
<p><u> Minipreps</u>
-
Following the standard  <a href=https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot4">protocol fot the miniprep  </a></br>
+
Following the standard  <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot4">protocol fot the miniprep  </a></br>
I measured DNA content with the nanodrop. </br>
I measured DNA content with the nanodrop. </br>
<u>Digestion analysis</u>: (Clovis's recipe)</br>
<u>Digestion analysis</u>: (Clovis's recipe)</br>
Line 335: Line 375:
<h5 id="date">July 11th</h5>
<h5 id="date">July 11th</h5>
<p><u> Minipreps</u> [yes, again!]
<p><u> Minipreps</u> [yes, again!]
-
Following the standard  <a href=https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot4">protocol fot the miniprep  </a></br>
+
Following the standard  <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot4">protocol fot the miniprep  </a></br>
I measured DNA content with the nanodrop. </br>
I measured DNA content with the nanodrop. </br>
<u>Digestion analysis</u>: (Clovis's recipe)</br>
<u>Digestion analysis</u>: (Clovis's recipe)</br>
Line 345: Line 385:
(Final volume of 50 uL)</br>
(Final volume of 50 uL)</br>
-
<u>Result:</u> The gel showed that I mislabelled the constructs. There were 3 promoters (big and very small fragments) </br> I will restart the experiment from the Biobricks
+
<u>Result:</u> The gel showed that I mislabelled the constructs. There were 3 promoters (big and very small fragments) </br> I will restart the experiment from the Biobricks</p>
 +
<p>Biobricks have been taken from the -20C stock and I followed <a href=kit>iGEM Distribution Kit instructions </a> and <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot1">Heat Shock transformation of <i>E.coli</i></a> </p>
 +
 +
</br>
-
</p>
+
<h5 id="date">July 12th</h5>
 +
<p>Colonies grew for all 3 Biobricks. </br> They have been put into a liquid culture in order to put them into the <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">Glycerol stock </a></p>
 +
</br>
-
</p>
+
<h5 id="date">July 13th</h5>
 +
<p> The <a href="https://2014.igem.org/Team:Paris_Bettencourt/Protocols#prot8">Glycerol stock </a> have been finish</p>

Revision as of 15:03, 21 August 2014

Paris Bettencourt 2014



Notebook

Interlab Study

July

July 4th

We will participate in the iGEM interlab study

Device 1

Existing device: BBa_I20260 (J23101-B0032-E0040-B0015) in the pSB3K3 vector.

    Kit location
  • Spring 2014 Plate 4, Well 18A

We use NEB Turbo cells to transform with the Biobrick

Biobrick extraction (iGEM protocol)

To use the DNA in the Distribution Kit you may follow these instructions:

  1. With a pipette tip, punch a hole through the foil cover into the corresponding well of the Biobrick™-standard part that you want.
  2. Pipette 10uL of dH2O (distilled water) into the well. Pipette up and down a few times and let sit for 5 minutes to make sure the dried DNA is fully resuspended.

Then we followed our protocol for Heat Shock transformation of E.coli

July 5th

Bacterias plated yesterday growed and made big and small colonies.I picked one small and one big colony from one plate in order to make liquid culture:

  1. 5mL of LB
  2. 5um Chlorophenicol
  3. Inoculate a single colony

July 6th

I made a glycerol stock from the liquid culture following the standard protocol of the Glycerol stock and labelled it with a number G.22



July 7th

Devices 2 and 3

New device to be built by the iGEM team: BBa_J23101 + BBa_E0240 (B0032-E0040-B0015), must be built in the pSB1C3 backbone

    Kit locations (Spring 2014)
  1. BBa_J23101 (called BBa_K823005 when in pSB1C3): Plate 1, Well 20K
  2. BBa_E0240 (in pSB1C3): Plate 2, Well 24B

New device to be built by the iGEM team: BBa_J23115 + BBa_E0240 (B0032-E0040-B0015), must be built in the pSB1C3 backbone
    Kit locations (Spring 2014)
  1. BBa_J23115 (called BBa_K823012 when in pSB1C3): Plate 1, Well 22I
  2. BBa_E0240 (in pSB1C3): Plate 2, Well 24B

I followed iGEM Distribution Kit instructions and Heat Shock transformation of E.coli


July 8th

    Successful transformations:
  • BBa_E0240
  • BBa_K823012

    Unsuccessful transformation:
  • BBa_K823005

For the successful transformations I followed the Glycerol stock protocol

For the unsuccessful transformation I transformed NEB turbo again with the rest of Biobrick BBa_K823005 following again iGEM Distribution Kit instructions and Heat Shock transformation of E.coli


July 9th

I finished the Glycerol stock protocol and labelled G.23 for BBa_K823012 and G.24 for BBa_E0240

BBa_K823005 successfully transformed! I followed the Glycerol stock protocol

Minipreps
Following the standard protocol fot the miniprep
I measured DNA content with the nanodrop.
Digestion analysis: (Clovis's recipe)
- 5 ug plasmid (given 211ng of BBa_E0240 - 15uL // given 387ng of BBa_K823012 - 30uL
- 5 ul FD Buffer
- 2.5 uL SpeI + 2.5 uL PstI (BBa_K823012) / 2.5 uL XbeI + 2.5 uL PstI (BBa_E0240)
- complete with H2O
(Final volume of 50 uL)

Result:DNA wasn't cut. Possible causes:
- wrongly calculated volumes (BBa_K823012 shouldn't be 30uL)
- too short digestion time


July 10th

Finished the Glycerol stock protocol and labelled it G.25

Minipreps Following the standard protocol fot the miniprep
I measured DNA content with the nanodrop.
Digestion analysis: (Clovis's recipe)
- 5 ug plasmid
- 5 ul FD Buffer
- 2.5 uL SpeI + 2.5 uL PstI (BBa_K823012) / 2.5 uL XbeI + 2.5 uL PstI (BBa_E0240)
- complete with H2O
(Final volume of 50 uL)
In the final 30uL of the miniprep of BBa_K823005 I didn't have enough of DNA.


July 11th

Minipreps [yes, again!] Following the standard protocol fot the miniprep
I measured DNA content with the nanodrop.
Digestion analysis: (Clovis's recipe)
- 5 ug plasmid
- 5 ul FD Buff(given 211ng of BBa_E0240 - 21uL // given 144ng and 120ng of BBa_K823012 - 35uL // given 273ng of BBa_K823005 - 20uL
- 2.5 uL SpeI + 2.5 uL PstI (BBa_K823012 and BBa_K823005) / 2.5 uL XbeI + 2.5 uL PstI (BBa_E0240)
- complete with H2O
(Final volume of 50 uL)
Result: The gel showed that I mislabelled the constructs. There were 3 promoters (big and very small fragments)
I will restart the experiment from the Biobricks

Biobricks have been taken from the -20C stock and I followed iGEM Distribution Kit instructions and Heat Shock transformation of E.coli


July 12th

Colonies grew for all 3 Biobricks.
They have been put into a liquid culture in order to put them into the Glycerol stock


July 13th

The Glycerol stock have been finish


August

Date 1

Text

Text

Date 2

Text

Text


September

Date 1

Text

Text

Date 2

Text

Text


October

Date 1

Text

Text

Date 2

Text

Text


Retrieved from "http://2014.igem.org/Team:Paris_Bettencourt/Notebook/Interlab_Study"