Team:Freiburg Bioware/NoteBook/Labjournal

From 2010.igem.org

(Difference between revisions)

Revision as of 01:05, 23 October 2010

Get to know our virus

Our project starts with first sequence analysis.

Checking iGEM compatibility

Scanning for iGEM restriction sites:
• Vector plasmid
• Thymidine kinase
• Cytosindeaminase

First practical steps

We took a closer look to the theoretical DNA- and amino acid sequence of the ITRs, questioned the role of the β-globin intron, had a closer look to the CMV promoter and performed our first cloning attempt.

First fluorescing HT1080 cells

We conducted our first site-directed mutagenesis.

First cell culture steps:
• Splitting and seeding of AAV 293 and HT1080 cells
• Calcium phosphate transfection
• Transduction with virus particles containing YFP encoding sequence
• Microscopy: Fluorescent transduced HT1080 cells

We planned to determine infectious virus titer via quantitative real-time PCR. For this purpose primers were designed, transduced HT1080 cells were harvested and prepared.

Theoretical studies of the AAV structure: Impressions.

Headline

https://2010.igem.org/Team:Freiburg_Bioware/NoteBook => Back to Notebook overview]

@@ Line 28: / Line 28: @@
 <div class="lab_journal_box">
 <div class="lab_journal_text">
-<br><h2>Headline</h2>
+<br><h2>Get to know our virus</h2>
-<!---Insert Text in here--->
+<!---Insert Text in here---> Our project starts with first sequence analysis.
 </div>
 <div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/March" ><img src="https://static.igem.org/mediawiki/2010/2/21/Freiburg10_Labjournal-March.png" /></a></div></div>
@@ Line 35: / Line 35: @@
 <div class="lab_journal_box">
 <div class="lab_journal_text">
-<br><h2>Headline</h2>
+<br><h2>Checking iGEM compatibility </h2>
-<!---Insert Text in here--->
+<!---Insert Text in here--->Scanning for iGEM restriction sites:<br/>
+•	Vector plasmid <br/>
+•	Thymidine kinase<br/>
+•	Cytosindeaminase<br/>
 </div>
 <div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/April" ><img src="https://static.igem.org/mediawiki/2010/6/6b/Freiburg10_Labjournal-April.png" /></a></div></div>
@@ Line 42: / Line 46: @@
 <div class="lab_journal_box">
 <div class="lab_journal_text">
-<br><h2>Headline</h2>
+<br><h2>First practical steps</h2>
-<!---Insert Text in here--->
+<!---Insert Text in here--->We took a closer look to the theoretical DNA- and amino acid sequence of the ITRs, questioned the role of the β-globin intron, had a closer look to the CMV promoter and performed our first cloning attempt.
 </div>
 <div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/May" ><img src="https://static.igem.org/mediawiki/2010/6/61/Freiburg10_Labjournal-May.png" /></a></div></div>
@@ Line 49: / Line 53: @@
 <div class="lab_journal_box">
 <div class="lab_journal_text">
-<br><h2>Headline</h2>
+<br><h2>First fluorescing HT1080 cells</h2>
-<!---Insert Text in here--->
+<!---Insert Text in here--->We conducted our first site-directed mutagenesis. <br/>
+<br/>
+First cell culture steps:<br/>
+•	Splitting and seeding of AAV 293 and HT1080 cells<br/>
+•	Calcium phosphate transfection<br/>
+•       Transduction with virus particles containing YFP encoding sequence<br/>
+•	Microscopy: Fluorescent transduced HT1080 cells<br/>
+<br/>
+We planned to determine infectious virus titer via quantitative real-time PCR. For this purpose primers were designed, transduced HT1080 cells were harvested and prepared.<br/>
+<br/>
+Theoretical studies of the AAV structure: Impressions.
 </div>
 <div class="lab_journal_img"><a href="https://2010.igem.org/Team:Freiburg_Bioware/NoteBook/Labjournal/June" ><img src="https://static.igem.org/mediawiki/2010/4/48/Freiburg10_Labjournal-June.png" /></a></div></div>