Team:JUIT-India/Notebook

From 2012.igem.org

(Difference between revisions)
(Prototype team page)
Line 1: Line 1:
-
<!-- *** What falls between these lines is the Alert Box!  You can remove it from your pages once you have read and understood the alert *** -->
 
-
 
<html>
<html>
-
<div id="box" style="width: 700px; margin-left: 137px; padding: 5px; border: 3px solid #000; background-color: #fe2b33;">
+
<style>
-
<div id="template" style="text-align: center; font-weight: bold; font-size: large; color: #f6f6f6; padding: 5px;">
+
#contentSub, #search-controls, .firstHeading, #footer-box, #catlinks, #p-logo {
-
This is a template page. READ THESE INSTRUCTIONS.
+
    display:none;}
-
</div>
+
#top-section {
-
<div id="instructions" style="text-align: center; font-weight: normal; font-size: small; color: #f6f6f6; padding: 5px;">
+
    border: 0px;
-
You are provided with this team page template with which to start the iGEM seasonYou may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki. You can find some examples <a href="https://2008.igem.org/Help:Template/Examples">HERE</a>.
+
    height: 0px;}
-
</div>
+
#content {
-
<div id="warning" style="text-align: center; font-weight: bold; font-size: small; color: #f6f6f6; padding: 5px;">
+
    border: none;}
-
You <strong>MUST</strong> have all of the pages listed in the menu below with the names specified. PLEASE keep all of your pages within your teams namespace.
+
/* Removes "teams" from the menubar */
-
</div>
+
#menubar > ul > li:last-child {
 +
    display: none;}
 +
/* Resizes the menubar to fik the links (default is 400px) */
 +
#menubar {
 +
    width: auto;}
 +
body {
 +
    margin: 10px 0 0 0;
 +
    padding: 5;}
 +
#top-section {
 +
    width: 965px;
 +
    height: 0;
 +
    margin: 0 auto;
 +
    padding: 0;
 +
    border: none;}
 +
#menubar {
 +
    font-size: 65%;
 +
    top: -14 px;}
 +
.left-menu:hover {
 +
    background-color: transparent;}
 +
#menubar li a {
 +
    background-color: transparent;}
 +
#menubar:hover {
 +
    color: white;}
 +
#menubar li a {
 +
    color: transparent;}
 +
#menubar:hover li a {
 +
    color: white;}
 +
 
 +
 
 +
img
 +
{
 +
opacity:1.0;
 +
filter:alpha(opacity=100);
 +
}
 +
img:hover
 +
{
 +
opacity:1.0;
 +
filter:alpha(opacity=100);
 +
}
 +
.cssmenu{
 +
border:none;
 +
border:0px;
 +
margin:0px;
 +
padding:0px;
 +
font: 67.5% 'Lucida Sans Unicode', 'Bitstream Vera Sans', 'Trebuchet Unicode MS', 'Lucida Grande', Verdana, Helvetica, sans-serif;
 +
font-size:14px;
 +
font-weight:bold;
 +
}
 +
.cssmenu ul{
 +
background:#333000;
 +
height:35px;
 +
list-style:none;
 +
margin:0;
 +
padding:0;
 +
}
 +
.cssmenu li{
 +
float:left;
 +
padding: 0px;
 +
}
 +
.cssmenu li a{
 +
background:#333333 url('https://static.igem.org/mediawiki/2012/3/3f/Seperator.gif') bottom right no-repeat;
 +
color:#cccccc;
 +
display:block;
 +
font-weight:normal;
 +
line-height:35px;
 +
margin:0px;
 +
padding:0px 25px;
 +
text-align:center;
 +
text-decoration:none;
 +
}
 +
.cssmenu li a:hover, .cssmenu ul li:hover a{
 +
background: #2580a2 url('https://static.igem.org/mediawiki/2012/e/e6/Hover.gif') bottom center no-repeat;
 +
 +
text-decoration:none;
 +
}
 +
.cssmenu li ul{
 +
background:#333333;
 +
display:none;
 +
height:auto;
 +
padding:0px;
 +
margin:0px;
 +
border:0px;
 +
position:absolute;
 +
width:225px;
 +
z-index:200;
 +
/*top:1em;
 +
/*left:0;*/
 +
}
 +
.cssmenu li:hover ul{
 +
display:block;
 +
 +
}
 +
.cssmenu li li {
 +
background:url('https://static.igem.org/mediawiki/2012/3/3f/Sub_sep.gif') bottom left no-repeat;
 +
display:block;
 +
float:none;
 +
margin:0px;
 +
padding:0px;
 +
width:225px;
 +
}
 +
.cssmenu li:hover li a{
 +
background:none;
 +
 +
}
 +
.cssmenu li ul a{
 +
display:block;
 +
height:35px;
 +
font-size:12px;
 +
font-style:normal;
 +
margin:0px;
 +
padding:0px 10px 0px 15px;
 +
text-align:left;
 +
}
 +
.cssmenu li ul a:hover, .cssmenu li ul li:hover a{
 +
background:#2580a2 url('https://static.igem.org/mediawiki/2012/a/a8/Hover_sub.gif') center left no-repeat;
 +
border:0px;
 +
 +
text-decoration:none;
 +
}
 +
.cssmenu p{
 +
clear:left;
 +
}
 +
</style>
 +
<div class='cssmenu' z-index:5000>
 +
<div z-index:100><img src="https://static.igem.org/mediawiki/2012/2/20/Lg.jpg" width="965" height="100"></div>
 +
<ul>
 +
  <li><a href='https://2012.igem.org/Team:JUIT-India'><span>Home</span></a></li>
 +
  <li><a href='#'><span>Team</span></a>
 +
      <ul>
 +
        <li><a href='https://2012.igem.org/Team:JUIT-India/Team'><span> Team</span></a></li>
 +
                  <li><a href='https://2012.igem.org/Team:JUIT-India/Sponsors'><span>Sponsors</span></a></li>
 +
              </ul>
 +
  </li>
 +
  <li><a href='#'><span>Project</span></a>
 +
      <ul>
 +
        <li><a href='https://2012.igem.org/Team:JUIT-India/Project'><span>Overview</span></a></li>
 +
        <li><a href='https://2012.igem.org/Team:JUIT-India/Protocol'><span>Protocol</span></a></li>
 +
      </ul>
 +
  </li>
 +
  <li><a href='https://2012.igem.org/Team:JUIT-India/Parts'><span>Parts</span></a></li> 
 +
  <li><a href='https://2012.igem.org/Team:JUIT-India/Design'><span>Design</span></a></li>
 +
  <li><a href='https://2012.igem.org/Team:JUIT-India/Modeling'><span>Modeling</span></a></li>
 +
  <li><a href='https://2012.igem.org/Team:JUIT-India/Human_Practices'><span>Human Practices</span></a>
 +
      <ul>
 +
          </a></li>
 +
 
 +
      </ul>
 +
  </li>
 +
  <li><a href='https://2012.igem.org/Team:JUIT-India/Notebook'><span>Notebook</span></a>
 +
  <ul>
 +
        <li><a href='https://2012.igem.org/Team:JUIT-India/Experiment'><span>Experimental Design</span></a></li>
 +
        <li><a href='https://2012.igem.org/Team:JUIT-India/WetLabWork'><span>Diary</span></a></li>
 +
        <li><a href='https://2012.igem.org/Team:JUIT-India/References'><span>Brainstorming</span></a></li>
 +
     
 +
      </ul>
 +
  </li>
 +
  <li><a href='https://2012.igem.org/Team:JUIT-India/Safety'><span>Safety</span></a></li>
 +
 
 +
</ul>
</div>
</div>
 +
<div z-index:100><img src="https://static.igem.org/mediawiki/2012/5/5d/Diary.jpg" width="965" height="300"></div>
 +
 +
<r>
 +
<hr>
 +
 +
 +
 +
 +
</html>
</html>
 +
Diary:
-
<!-- *** End of the alert box *** -->
+
== ''' June ''' ==
-
{| style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" bordercolor="#fff" width="62%" align="center"
+
• Brainstorming various ideas
-
!align="center"|[[Team:JUIT-India|Home]]
+
<br>
-
!align="center"|[[Team:JUIT-India/Team|Team]]
+
• Decide the project to work on for this year
-
!align="center"|[https://igem.org/Team.cgi?year=2012&team_name=JUIT-India Official Team Profile]
+
-
!align="center"|[[Team:JUIT-India/Project|Project]]
+
-
!align="center"|[[Team:JUIT-India/Parts|Parts Submitted to the Registry]]
+
-
!align="center"|[[Team:JUIT-India/Modeling|Modeling]]
+
-
!align="center"|[[Team:JUIT-India/Notebook|Notebook]]
+
-
!align="center"|[[Team:JUIT-India/Safety|Safety]]
+
-
!align="center"|[[Team:JUIT-India/Attributions|Attributions]]
+
-
|}
+
 +
 +
==July:==
 +
<br>• Week 1
 +
<br>o Research on the project thoroughly
 +
<br>o Primers Designing
 +
• Week 2
 +
<br>o Culturing of M.Capsulatus
 +
<br>o Isolation of Genomic DNA from M.Capsulatus
 +
<br>o PCR reaction for amplification of MxaF
 +
<br>o Agarose Gel Electrophoresis
 +
<br>o Optimization & Standardization of PCR
 +
<br> Monday : Cultured M.Capsulatus
 +
<br> Tuesday : Isolated Genomic DNA from M.Capsulatus
 +
<br>Agarose Gel Electrophoresis
 +
<br> Wednesday : PCR Reaction  for amplification of MxaF
 +
<br>Agarose Gel Electrophoresis
 +
Result: Non-Specific Binding<br>
 +
 Thursday : PCR reaction performed again
 +
<br>Agarose Gel Electrophoresis<br>
 +
Result: NO bands were observed
 +
<br> Friday : PCR reaction performed
 +
<br>Agarose Gel Electrophoresis<br>
 +
Result: NO bands were observed
 +
<br> Saturday: PCR reaction performed
 +
Agarose Gel Electrophoresis<br>
 +
Result: light bands were observed
 +
<br> Sunday: PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: bands were observed
 +
<br>Gel Extraction.
 +
 +
 +
<br>• Week 3
 +
<br>o Culturing of M.Capsulatus
 +
<br>o Isolation of Genomic DNA from M.Capsulatus
 +
<br>o PCR reaction for amplification of NifA
 +
<br>o Agarose Gel Electrophoresis
 +
<br>o Optimization & Standardization of PCR
 +
<br> Monday : Cultured M.Capsulatus
 +
<br> Tuesday : Isolated Genomic DNA from M.Capsulatus
 +
<br>Agarose Gel Electrophoresis
 +
<br>Results: NO band was observed (due to less duration in the -200c freezer
 +
<br> Wednesday : Isolated Genomic DNA from M.Capsulatus
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: Sharp Bands were observed
 +
<br> Thursday : PCR reaction performed for amplification of NifA
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: NO bands were observed
 +
<br> Friday : PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: Nonspecific bands were observed
 +
<br> Saturday: PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: Nonspecific bands were observed
 +
<br> Sunday: PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: Sharp bands were observed
 +
<br>Gel Extraction.
 +
 +
<br>• Week 4
 +
<br>o Culturing of B.subtilis
 +
<br>o Isolation of genomic DNA from B.subtilis
 +
<br>o PCR reaction for amplification of SacB
 +
<br>o Agarose Gel Electrophoresis
 +
<br>o Optimization & Standardization of PCR
 +
<br> Monday : Cultured B.subtilis
 +
<br> Tuesday : Isolated Genomic DNA from B.subtilis
 +
<br>Agarose Gel Electrophoresis
 +
<br> Wednesday : PCR Reaction  for amplification of SacB
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: Non-Specific Binding
 +
<br> Thursday : PCR reaction performed again
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: NO bands were observed
 +
<br> Friday : PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: NO bands were observed
 +
<br> Saturday: PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: light bands were observed
 +
<br> Sunday: PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
<br>Result: bands were observed
 +
<br>Gel Extraction was performed.
 +
 +
 +
==August: ==
 +
<br>• Week 1
 +
<br>o Culturing of P. aeruginosa
 +
<br>o Isolation of genomic DNA from P. aeruginosa
 +
<br>o PCR Reaction for amplification of NosZ
 +
<br>o Agarose Gel Electrophoresis
 +
<br>o Optimization & Standardization of PCR
 +
<br> Monday : Cultured P. aeruginosa
 +
<br> Tuesday : Isolated Genomic DNA from P. aeruginosa
 +
Agarose Gel Electrophoresis
 +
Results: NO band was observed
 +
<br> Wednesday : Isolated Genomic DNA from P. aeruginosa using CTAB method
 +
Agarose Gel Electrophoresis
 +
Result: Sharp Bands were observed
 +
<br> Thursday : PCR reaction performed for amplification of NosZ
 +
<br>Agarose Gel Electrophoresis
 +
Result: NO bands were observed
 +
<br> Friday : PCR reaction performed
 +
Agarose Gel Electrophoresis
 +
Result: No bands were observed
 +
<br> Saturday: PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
Result: Nonspecific bands were observed
 +
<br> Sunday: PCR reaction performed
 +
<br>Agarose Gel Electrophoresis
 +
Result: Sharp bands were observed
 +
<br>Gel Extraction was performed.
 +
 +
 +
 +
<br>• Week 2
 +
<br>o PCR reactions for amplification of MxaF & NifA
 +
<br>o Gel Extraction for the genes
 +
<br>o Ligation of genes into TA Vector
 +
<br>o Transformation of E.Coli cells with TA Vector
 +
<br>o Culturing of Transformed Cells –  Starter Culture
 +
<br> Monday : PCR reaction performed for MxaF & NifA
 +
<br>Agarose Gel Electrophoresis
 +
Result: Sharp bands were observed
 +
<br>Gel Extraction was performed.
 +
<br> Tuesday : Ligation of MxaF into TA Vector
 +
<br> Wednesday : Transformation using MgCl2 & CaCl2 method
 +
                        Result: Failed(Contamination)
 +
<br> Thursday : Ligation of MxaF into TA Vector
 +
<br> Friday:      Transformation using MgCl2 & CaCl2 method
 +
                        Result: White Colonies were selected and cultured
 +
<br> Saturday: Ligation of NifA into TA Vector
 +
<br> Sunday: :      Transformation using MgCl2 & CaCl2 method
 +
                        Result: White Colonies were selected and cultured
 +
o
 +
 +
<br>• Week 3
 +
<br>o PCR reactions for amplification of NosZ & SacB
 +
<br>o Gel Extraction for the genes
 +
<br>o Ligation of genes into TA Vector
 +
<br>o Transformation of E.Coli cells with TA Vector
 +
<br>o Culturing of Transformed Cells – Starter Culture\
 +
<br> Monday : PCR reaction performed for NosZ & SacB
 +
<br>Agarose Gel Electrophoresis
 +
Result: Sharp bands were observed
 +
<br>Gel Extraction was performed.
 +
<br> Tuesday : Ligation of NosZ into TA Vector<br>
 +
 Wednesday : Transformation using MgCl2 & CaCl2 method
 +
 Result: White Colonies were selected and cultured
 +
 Thursday : Ligation of SacB into TA Vector<br>
 +
 Friday:      Transformation using MgCl2 & CaCl2 method<br>
 +
                        Result: No colonies were observed<br>
 +
 Saturday: Ligation of SacB into TA Vector <br>
 +
 Sunday: :      Transformation using MgCl2 & CaCl2 method<br>
 +
                        Result: White Colonies were selected and cultured<br>
 +
 +
• Week 4<br>
 +
o Culture transformed Cells<br>
 +
o Plasmid Isolation<br>
 +
o PCR reactions to confirm the insertion of genes<br>
 +
o Agarose Gel Electrophoresis<br>
 +
 Monday : Cultured MxaF, NifA, SacB & NosZ transformed Cells<br>
 +
 Tuesday : Plasmid Isolation for MxaF & NifA transformed cells
 +
                  Restriction Digestion of the plasmids<br>
 +
 Wednesday : PCR reaction performed for amplification of MxaF & NifA
 +
                    Agarose Gel Electrophoresis.  <br>
 +
                    Results: Sharp Bands were observed.<br>
 +
 Thursday : Plasmid Isolation for NosZ & SacB transformed cells
 +
                  Restriction Digestion of the plasmids.<br>
 +
                  Agarose Gel Electrophoresis<br>
 +
 Friday:      PCR reaction performed for amplification of MxaF & NifA <br>
 +
 Saturday:  Agarose Gel Electrophoresis<br>
 +
 +
 +
==September: ==
 +
• Week 1<br>
 +
o Restriction Digestion of MxaF containing plasmids<br>
 +
o Ligation of MxaF into psb1c3<br>
 +
o Restriction Digestion of NosZcontaining plasmids<br>
 +
o Ligation of NosZ into psb1c3 containg MxaF<br>
 +
 Monday : Restriction Digestion of MxaF containing plasmids<br>
 +
Agarose Gel Electrophoresis<br>
 +
Result: Sharp bands were observed<br>
 +
Gel Extraction was performed.<br>
 +
 Tuesday : Restriction Digestion of psb1c3 using EcoR1
 +
                Ligation of MxaF into psb1c3<br>
 +
 Thursday : Restriction Digestion of NosZ containing plasmids<br>
 +
Agarose Gel Electrophoresis<br>
 +
Result: Sharp bands were observed<br>
 +
 Friday:      Transformation using MgCl2 & CaCl2 method      <br>             
 +
 Saturday: Restriction Digestion of psb1c3<br>
 +
                  Ligation of NosZ into psb1c3 containing MxaF<br>
 +
 +
• <b>Week 2:</b><br>
 +
o Restriction Digestion of NifA<br>
 +
o Ligation of NifA into psb1c3 containg NifA & SacB<br>
 +
o Restriction Digestion of SacB<br>
 +
o Ligation of SacB into psb1c3 containg all the other genes<br>
 +
 Monday : Restriction Digestion of NifA containing plasmids<br>
 +
Agarose Gel Electrophoresis<br>
 +
Result: Sharp bands were observed<br>
 +
Gel Extraction was performed.<br>
 +
 Tuesday : Restriction Digestion of psb1c3 containing MxaF & NosZ using EcoR1
 +
                Ligation of NifA into psb1c3<br>
 +
 Thursday : Restriction Digestion of SacB containing plasmids<br>
 +
Agarose Gel Electrophoresis<br>
 +
Result: Sharp bands were observed<br>
 +
 Friday:      Transformation using MgCl2 & CaCl2 method                    <br>
 +
 Saturday: Restriction Digestion of psb1c3<br>
 +
                  Ligation of SacB into psb1c3 containing all the other genes.<br>
 +
 +
• <b>Week 3:</b><br>
 +
o Transformation of M.Capsulatus competent Cells using the plasmid containing all the genes<br>
 +
o Growth of transformed cells on selective media.<br>
 +
o Plasmid Isolation <br>
 +
o PCR reaction to confirm the insertion of genes<br>
 +
 Monday: Cultures M.Capsulatus Cells<br>
 +
 Tuesday : Preparation of M.Capsulatus Competent Cells<br>
 +
 Wednesday: Transformation of competent cells using the prepared plasmids<br>
 +
 Thursday : Growth of plates on LB plates containg chloramphenicol<br>
 +
 Friday:      Cultured transformed cells in LB agar<br>
 +
 Saturday: Plasmid Isolation <br>
 +
                  Restriction Digestion<br>
 +
 Sunday: Agarose Gel Electrophoresis<br>
 +
 +
 +
 +
 +
== Sponsors ==
 +
Our Title Sponsor :
 +
<html>
 +
<br>
 +
<center>
 +
<img src="https://static.igem.org/mediawiki/2012/2/2c/Juit_logo.png" >
 +
</center>
 +
</html>
-
You should make use of the calendar feature on the wiki and start a lab notebook.  This may be looked at by the judges to see how your work progressed throughout the summer.  It is a very useful organizational tool as well.
+
{|align="justify"

Revision as of 09:38, 26 September 2012


Diary:

Contents

June

• Brainstorming various ideas
• Decide the project to work on for this year


July:


• Week 1
o Research on the project thoroughly
o Primers Designing • Week 2
o Culturing of M.Capsulatus
o Isolation of Genomic DNA from M.Capsulatus
o PCR reaction for amplification of MxaF
o Agarose Gel Electrophoresis
o Optimization & Standardization of PCR
 Monday : Cultured M.Capsulatus
 Tuesday : Isolated Genomic DNA from M.Capsulatus
Agarose Gel Electrophoresis
 Wednesday : PCR Reaction for amplification of MxaF
Agarose Gel Electrophoresis Result: Non-Specific Binding
 Thursday : PCR reaction performed again
Agarose Gel Electrophoresis
Result: NO bands were observed
 Friday : PCR reaction performed
Agarose Gel Electrophoresis
Result: NO bands were observed
 Saturday: PCR reaction performed Agarose Gel Electrophoresis
Result: light bands were observed
 Sunday: PCR reaction performed
Agarose Gel Electrophoresis
Result: bands were observed
Gel Extraction.



• Week 3
o Culturing of M.Capsulatus
o Isolation of Genomic DNA from M.Capsulatus
o PCR reaction for amplification of NifA
o Agarose Gel Electrophoresis
o Optimization & Standardization of PCR
 Monday : Cultured M.Capsulatus
 Tuesday : Isolated Genomic DNA from M.Capsulatus
Agarose Gel Electrophoresis
Results: NO band was observed (due to less duration in the -200c freezer
 Wednesday : Isolated Genomic DNA from M.Capsulatus
Agarose Gel Electrophoresis
Result: Sharp Bands were observed
 Thursday : PCR reaction performed for amplification of NifA
Agarose Gel Electrophoresis
Result: NO bands were observed
 Friday : PCR reaction performed
Agarose Gel Electrophoresis
Result: Nonspecific bands were observed
 Saturday: PCR reaction performed
Agarose Gel Electrophoresis
Result: Nonspecific bands were observed
 Sunday: PCR reaction performed
Agarose Gel Electrophoresis
Result: Sharp bands were observed
Gel Extraction.


• Week 4
o Culturing of B.subtilis
o Isolation of genomic DNA from B.subtilis
o PCR reaction for amplification of SacB
o Agarose Gel Electrophoresis
o Optimization & Standardization of PCR
 Monday : Cultured B.subtilis
 Tuesday : Isolated Genomic DNA from B.subtilis
Agarose Gel Electrophoresis
 Wednesday : PCR Reaction for amplification of SacB
Agarose Gel Electrophoresis
Result: Non-Specific Binding
 Thursday : PCR reaction performed again
Agarose Gel Electrophoresis
Result: NO bands were observed
 Friday : PCR reaction performed
Agarose Gel Electrophoresis
Result: NO bands were observed
 Saturday: PCR reaction performed
Agarose Gel Electrophoresis
Result: light bands were observed
 Sunday: PCR reaction performed
Agarose Gel Electrophoresis
Result: bands were observed
Gel Extraction was performed.


August:


• Week 1
o Culturing of P. aeruginosa
o Isolation of genomic DNA from P. aeruginosa
o PCR Reaction for amplification of NosZ
o Agarose Gel Electrophoresis
o Optimization & Standardization of PCR
 Monday : Cultured P. aeruginosa
 Tuesday : Isolated Genomic DNA from P. aeruginosa Agarose Gel Electrophoresis Results: NO band was observed
 Wednesday : Isolated Genomic DNA from P. aeruginosa using CTAB method Agarose Gel Electrophoresis Result: Sharp Bands were observed
 Thursday : PCR reaction performed for amplification of NosZ
Agarose Gel Electrophoresis Result: NO bands were observed
 Friday : PCR reaction performed Agarose Gel Electrophoresis Result: No bands were observed
 Saturday: PCR reaction performed
Agarose Gel Electrophoresis Result: Nonspecific bands were observed
 Sunday: PCR reaction performed
Agarose Gel Electrophoresis Result: Sharp bands were observed
Gel Extraction was performed.



• Week 2
o PCR reactions for amplification of MxaF & NifA
o Gel Extraction for the genes
o Ligation of genes into TA Vector
o Transformation of E.Coli cells with TA Vector
o Culturing of Transformed Cells – Starter Culture
 Monday : PCR reaction performed for MxaF & NifA
Agarose Gel Electrophoresis Result: Sharp bands were observed
Gel Extraction was performed.
 Tuesday : Ligation of MxaF into TA Vector
 Wednesday : Transformation using MgCl2 & CaCl2 method

                       Result: Failed(Contamination)


 Thursday : Ligation of MxaF into TA Vector
 Friday: Transformation using MgCl2 & CaCl2 method

                       Result: White Colonies were selected and cultured


 Saturday: Ligation of NifA into TA Vector
 Sunday: : Transformation using MgCl2 & CaCl2 method

                       Result: White Colonies were selected and cultured

o


• Week 3
o PCR reactions for amplification of NosZ & SacB
o Gel Extraction for the genes
o Ligation of genes into TA Vector
o Transformation of E.Coli cells with TA Vector
o Culturing of Transformed Cells – Starter Culture\
 Monday : PCR reaction performed for NosZ & SacB
Agarose Gel Electrophoresis Result: Sharp bands were observed
Gel Extraction was performed.
 Tuesday : Ligation of NosZ into TA Vector
 Wednesday : Transformation using MgCl2 & CaCl2 method  Result: White Colonies were selected and cultured  Thursday : Ligation of SacB into TA Vector
 Friday: Transformation using MgCl2 & CaCl2 method

                       Result: No colonies were observed

 Saturday: Ligation of SacB into TA Vector
 Sunday: : Transformation using MgCl2 & CaCl2 method

                       Result: White Colonies were selected and cultured

• Week 4
o Culture transformed Cells
o Plasmid Isolation
o PCR reactions to confirm the insertion of genes
o Agarose Gel Electrophoresis
 Monday : Cultured MxaF, NifA, SacB & NosZ transformed Cells
 Tuesday : Plasmid Isolation for MxaF & NifA transformed cells

                 Restriction Digestion of the plasmids

 Wednesday : PCR reaction performed for amplification of MxaF & NifA

                    Agarose Gel Electrophoresis.   
Results: Sharp Bands were observed.

 Thursday : Plasmid Isolation for NosZ & SacB transformed cells

                 Restriction Digestion of the plasmids.
Agarose Gel Electrophoresis

 Friday: PCR reaction performed for amplification of MxaF & NifA
 Saturday: Agarose Gel Electrophoresis


September:

• Week 1
o Restriction Digestion of MxaF containing plasmids
o Ligation of MxaF into psb1c3
o Restriction Digestion of NosZcontaining plasmids
o Ligation of NosZ into psb1c3 containg MxaF
 Monday : Restriction Digestion of MxaF containing plasmids
Agarose Gel Electrophoresis
Result: Sharp bands were observed
Gel Extraction was performed.
 Tuesday : Restriction Digestion of psb1c3 using EcoR1

                Ligation of MxaF into psb1c3

 Thursday : Restriction Digestion of NosZ containing plasmids
Agarose Gel Electrophoresis
Result: Sharp bands were observed
 Friday: Transformation using MgCl2 & CaCl2 method
 Saturday: Restriction Digestion of psb1c3

                  Ligation of NosZ into psb1c3 containing MxaF

Week 2:
o Restriction Digestion of NifA
o Ligation of NifA into psb1c3 containg NifA & SacB
o Restriction Digestion of SacB
o Ligation of SacB into psb1c3 containg all the other genes
 Monday : Restriction Digestion of NifA containing plasmids
Agarose Gel Electrophoresis
Result: Sharp bands were observed
Gel Extraction was performed.
 Tuesday : Restriction Digestion of psb1c3 containing MxaF & NosZ using EcoR1

                Ligation of NifA into psb1c3

 Thursday : Restriction Digestion of SacB containing plasmids
Agarose Gel Electrophoresis
Result: Sharp bands were observed
 Friday: Transformation using MgCl2 & CaCl2 method
 Saturday: Restriction Digestion of psb1c3

                  Ligation of SacB into psb1c3 containing all the other genes.

Week 3:
o Transformation of M.Capsulatus competent Cells using the plasmid containing all the genes
o Growth of transformed cells on selective media.
o Plasmid Isolation
o PCR reaction to confirm the insertion of genes
 Monday: Cultures M.Capsulatus Cells
 Tuesday : Preparation of M.Capsulatus Competent Cells
 Wednesday: Transformation of competent cells using the prepared plasmids
 Thursday : Growth of plates on LB plates containg chloramphenicol
 Friday: Cultured transformed cells in LB agar
 Saturday: Plasmid Isolation

                  Restriction Digestion

 Sunday: Agarose Gel Electrophoresis



Sponsors

Our Title Sponsor :