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Latest revision as of 23:13, 27 September 2013

FA Quantification and Cell Viability Module's Notebook

June 2013

Week 4

June 25:

Prepared DMEM in TC room
Added FBS (Serum) & Antibiotics into DMEM

June 27:

Designed the experiment for testing FA concentration by GCMS

June 28:

HepG2 cell culturing (first generation)

July 2013

Week 1

July 2:

HepG2 cell passaging
Made sodium palmitate from powder to solution by 50% Ethanol

July 3:

Used GCMS to quantify the FA in the medium

July 4:

Passaged HepG2 cells
Changed medium for cell line
Searched for information of MTT assay and Transfection by lipofectamine
Got GCMS results for the last day: Not accurate/usable

July 5:

Repeated GCMS test again testing FA in medium and we made
Got the result of GCMS: Not accurate/usable

Week 2

July 8:

Passaged HepG2 cells
Used some HepG2 cells to extract genomic DNA

July 9:

Another try of GCMS with a gradient concentration of FA
Got the result of GCMS: Not match with the concentration we made

July 10:

Repeated GCMS: No accurate results
Changed medium for the cell line
Checked lipofectamine protocol

July 11:

GCMS again for the same sodium palmitate we made: No accurate results
Passaged HepG2 cell line

July 12:

Prepared reagents for MTT assay for cell viability test, protocols checked
Changed the medium of the cell line

Week 3

July 15:

Seeded HepG2 cells into 96-well plate for MTT cell viability test and adding FA
Seeded HepG2 cells into 96-well plate for transfection of PEGFP-N1

July 16:

Transfection of PEGFP-N1 into HepG2 cells
Passaged HepG2 cells

July 17:

Changed the medium after transfection had been done in 24 hours
Did MTT assay and getting result (adding FA in the wrong time, cannot be used)
Made new DMEM
Changed medium for the cell line

July 18:

Seeded HepG2 cells in 24-well plate for transfection of PEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP
Got polylysine for coating

July 19:

Changed the medium for the HepG2 cell line
Seeded cells for another MTT assay
Checked HepG2 cells for transfection: growing slow, 30% confluency, cannot do transfection
Seeded cells on a 96-well plate for transfection in the coming week
Passaged HepG2 cells

Week 4

July 22:

Transfection of PEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP on 96-well plate
Added gradient concentration of sodium palmitate into HepG2 cells for MTT assay
Coated coverslips with polylysine for HepG2 cell staining and fixation

July 23:

Replaced the medium after transfection of PEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP
Finished MTT assay to get cell viability in different FA concentrations in 24 hours
Transferred cells containing PEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP into 24-well plate with well-coated coverslips in each well

July 24:

Stained mitochondria by Mito-tracker@Red and fixating by formaldehyde of HepG2 cells transfected with pEGFP-N1, pCMV/myc/mito/EGFP and Pmv/myc/mito/GFP in 24-well plate
Passaged HepG2 cells and changed medium of the cell line

July 25:

Got results for cell staining and fixation: No obvious overlapping pattern between GPF signal and Mito-tracker red.

July 26:

Coated coverslips by polylysine
Seeded HepG2 cells in one 24-well plate for transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP
Seeded HepG2 cells on two 96-well plate, for transfection of FABP construct and MTT assay in 48 hours

Week 5

July 29:

Transfection of FABP construct with pEGFP-N1 as control
Transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP in 24-well plate with well-coated coverslips
Added gradient concentration form 0.1mM to 1.2mM of FA into the plate for MTT assay

July 30:

Changed medium after transfection of pEGFP-N1 and FABP construct and saw the results: Both pEGFP-N1 and FABP construct did not show GFP signals
Changed medium after transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP
Did staining and fixation of the coverslips and saw the results: No obvious overlapping patterns of GFP signals and Mito-tracker red
Passaged HepG2 cells

August 2013

Week 1

Aug 1:

Seeded cells for transfection of pEGFP-N1, FABP construct, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP
Changed medium for HepG2 cell line
Got results for MTT assay in 48 hours

Aug 2:

Checked if transfection (after 22 hrs) worked

No GFP signal for pEFGP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP

Week 2

Aug 5:

Transfection of HepG2 cells on 10cm Plates
Passaged P(n+8) HepG2 cells into one P(n+9)
Seeded HepG2 cells on 96-well plates for transfection

Aug 6:

Trouble shooting for transfection of HepG2 cells on 96 well plates

Aug 7:

Observe transfected cell. GFP signal expressed
Seeded cells for transfection of pEGFP-N1, and (FABP construct/EGFP) in 96 well-plate
Seeded cells for transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP in 24 well plate with polylysine coated coverslips
Seeded cells for MTT assay

Aug 8:

Observed transfection of pEGFP-N1, and (FABP construct/EGFP) in 96 well-plate. No GFP signal observed
Observed transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP. GFP signal observed
Stained mitochondria of cells transfected with pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP using mito tracking dye
Observed cell with both transfection and stained mitochondria: No GFP signal observed because transfection occurred with very low efficiency.
Prepared new cell line: HEK293FT
Conducted MTT assay

Week 3

Aug 12:

MTT Assay without seeding and incubation
Changed medium for HepG2 and HEK293FT cell lines
Prepared polylysine coated coverslips

Aug 13:

Seeded cells using HEK293FT cell line for transfection

Aug 15:

Cells detached and not in good condition for transfection

Aug 16:

Seeded cells HEK293FT cell line for transfection

Week 4

Aug 19:

Transfection of pEGFP-N1, and (FABP construct/EGFP) in 96 well plate and pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP in 24 well plate.

Aug 20:

Seeing results of transfection of FABP construct and pEGFP-N1 in HEK293FT cells and seeing the results: pEGFP-N1 had signal but FABP did not show GFP signals

Aug 22:

Transfection of pEGFP-N1 and FABP construct into HEK293FT cells
Changing medium for HEK293FT cells and HepG2 cells and seeing the results: pEGFP-N1 had signal but FABP did not show GFP signals

Week 5

Aug 26:

Transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP in 60mm TC plates
Changing medium of transfection of the previous day and seeing the results: pEGFP-N1 had signal but FABP did not show GFP signals

Aug 28:

Changing medium of transfection of the previous day
Staining and fixation of
Drawing MTT standard curves

Aug 29:

Transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP into HEK293FT cells
Maintaining HEK293FT and HepG2 cell line

Aug 30:

Staining and fixation for HEK cells transfected by pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP
Seeing the results for transfected HEK293FT cells: No obvious overlapping pattern of GFP signals and Mito-tracker
Seeding HepG2 cells for Transfection of FABP construct and pEGFP-N1 in 96-well plate
Seeding HEK 293FT cells for transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP
Passaging HEK293FT cell line

Aug 31:

Transfection with pEGFP-N1 and pCMV/PolyA/EGFP into HEK293FT cells

September 2013

Week 1

Sept 1:

Checking results for transfection on Aug 31: pEGFP-N1 had strong signal while PolyA did not have signals

Sept 2:

Transfection of FABP construct and pEGFP-N1 in HepG2 cells
Check results for HEK293FT cells been transfected by pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP, by using confocal microscope

Sept 3:

Adding FA into HepG2 cells been transfected by FABP construct and pEGFP-N1
Maintaining HepG2 and HEK293FT cell line
Drug selection Test on HEK293FT test by purmycin

Sept 4:

Drug selection test on HEK293FT cells by neomycin and puromycin

Sept 5:

Staining and fixation of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP and seeing he results by confocal microscope
Making mew DMEM
Drug selection test observation

Sept 6:

Transfection of FAPB and pEGFP-N1 into HepG2 cells in 60 mm plates
Maintaining cell line for HEK293FT cells

Sept 7:

Checking results for transfection on Sept 6: pEGFP-N1 showed weak GFP signal, no signals for FABP
Change medium for drug selection test

Sept 8:

Maintaining cell line for HEK293FT cells and HepG2 cells

Week 2

Sept 9:

Checking transfection for FABP construct again: Weak signals were shown from the debris of the cells
Transfection of AceA construct

Sept 11:

Seeding HEK293FT cells into 35mm plates for transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP
Cell splitting of ACE

Sept 12:

Transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP into HEK293FT cells

Week 3

Content 1

Week 4

Content 1

Week 5

Content 1

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+ <a href="https://2013.igem.org/Main_Page"><img id="iGEM_Logo" src="https://static.igem.org/mediawiki/2013/4/46/Igem_qgem_logo.png"></a>
+<a href="http://www.ust.hk/eng/index.htm"><img id="hkust_Logo" src="https://static.igem.org/mediawiki/2013/5/55/Hkust_logo.gif"></a>
+<ol class="pos_fixed">
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/notebook">Notebook</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/notebook/mod1">Module 1</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/notebook/mod2">Module 2</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/notebook/mod3">Module 3</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/notebook/mod4">Module 4</a></il>
+</ol>
 <a href=https://2013.igem.org/Team:Hong_Kong_HKUST><center><div id="kepala" style="height:121px;width:100%;"><img src="https://static.igem.org/mediawiki/igem.org/c/c7/BANNER1_%281%29.png" style="height:121px;width:100%;align:middle;"></div></center></a>
 <div id="cover"></div>
@@ Line 254: / Line 314: @@
 <li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/advisors">Advisors</a></li>
 <li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/instructors">Instructors</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/attribution">Attribution</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/acknowledge">Acknowledgement</a></li>
 </ul>
 </li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/project">Project</a>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/Project">Project</a>
 <ul>
 <li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/abstract">Abstract</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/modelling">Modelling</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/modules">Modules Description</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/characterization">Characterization</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/data">Data Page</a></li>
 <li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/Parts">Parts</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/datapage">Data Page</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/characterization">Characterization</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/results">Results</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/results">Result</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/future">Future Work</a></li>
 </ul>
 </li>
@@ Line 270: / Line 333: @@
 <li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/Wetlab">Wetlab</a>
 <ul>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/notebook">notebook</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/notebook">Notebook</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/protocols">protocols</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/protocols">Protocols</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/safety">safety</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/safety">Safety</a></li>
 </ul>
 </li>
@@ Line 285: / Line 349: @@
 <li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp">Human Practice</a>
 <ul>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/Interviews">Interviews</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/cp">Country Profile</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/Presentation">Presentation</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/blog">Blog</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/Videos">Videos</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/interview">Interviews</a></li>
-<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/Article">Article</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/article/genet">Article</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/video">Videos</a></li>
+<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/hp/presentation">Presentations</a></li>
 </ul>
 </li>
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 </div>
-<br><br><br><br><br><br><br>
+<br><br><br><br><br><br><br><br>
+<div id="title"><center><h3>FA Quantification and Cell Viability Module's Notebook</h3></center></div>
 <div id="flight">
   <div id="satu" align="center"> <h1>June 2013</h1>
@@ Line 300: / Line 368: @@
        <div>
        <a href='#' class='head'>Week 4</a>
-       <div class='content' align="left">Wet lab<br>
+       <div class='content' align="left">
-·      Inoculation of pBlueScript KS(+) for training<br>
+<p>&nbsp;</p>
-·      Autoclave basic materials<Br>
+<p><strong>June 25:</strong></p>
-·      Preparing LB<br>
+<ul>
-·      LB-Ampicillin plates poured<br>
+  <li>Prepared DMEM in TC room</li>
-Dry lab<br>
+  <li>Added FBS (Serum) &amp; Antibiotics into DMEM</li>
-Protocols review
+</ul>
+<p>&nbsp;</p>
+<p><strong>June 27:</strong></p>
+<ul>
+  <li>Designed the experiment for testing FA concentration by GCMS</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>June 28:</strong></p>
+<ul>
+  <li>HepG2 cell culturing (first generation)</li>
+  <li></li>
+</ul><br>
        </div>
        </div>
@@ Line 316: / Line 395: @@
     <div>
        <a href='#' class='head'>Week 1</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>July 2:</strong></p>
+<ul>
+  <li>HepG2 cell passaging</li>
+  <li>Made sodium palmitate from powder to solution by 50% Ethanol</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 3:</strong></p>
+<ul>
+  <li>Used GCMS to quantify the FA in the medium</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 4:</strong></p>
+<ul>
+  <li>Passaged HepG2 cells</li>
+  <li>Changed medium for cell line</li>
+  <li>Searched for information of MTT assay and Transfection by lipofectamine</li>
+  <li>Got GCMS results for the last day: Not accurate/usable</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 5:</strong></p>
+<ul>
+  <li>Repeated GCMS test again testing FA in medium and we made</li>
+  <li>Got the result of GCMS: Not accurate/usable</li>
+</ul>
+<p>&nbsp;</p>
        </div>
     </div>
@@ Line 322: / Line 427: @@
     <div>
        <a href='#' class='head'>Week 2</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>July 8:</strong></p>
+<ul>
+  <li>Passaged HepG2 cells</li>
+  <li>Used some HepG2 cells to extract genomic DNA</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 9:</strong></p>
+<ul>
+  <li>Another try of GCMS with a gradient concentration of FA</li>
+  <li>Got the result of GCMS: Not match with the concentration we made</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 10:</strong></p>
+<ul>
+  <li>Repeated GCMS: No accurate results</li>
+  <li>Changed medium for the cell line</li>
+  <li>Checked lipofectamine protocol</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 11:</strong></p>
+<ul>
+  <li>GCMS again for the same sodium palmitate we made: No accurate results</li>
+  <li>Passaged HepG2 cell line</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 12:</strong></p>
+<ul>
+  <li>Prepared reagents for MTT assay for cell viability test, protocols checked</li>
+  <li>Changed the medium of the cell line</li>
+</ul>
+<p>&nbsp;</p>
        </div>
     </div>
@@ Line 328: / Line 465: @@
     <div>
        <a href='#' class='head'>Week 3</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>July 15:</strong></p>
+<ul>
+  <li>Seeded HepG2 cells into 96-well plate for MTT cell viability test and adding FA</li>
+  <li>Seeded HepG2 cells into 96-well plate for transfection of PEGFP-N1</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 16:</strong></p>
+<ul>
+  <li>Transfection of PEGFP-N1 into HepG2 cells</li>
+  <li>Passaged HepG2 cells</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 17:</strong></p>
+<ul>
+  <li>Changed the medium after transfection had been done in 24 hours</li>
+  <li>Did MTT assay and getting result (adding FA in the wrong time, cannot be used)</li>
+  <li>Made new DMEM</li>
+  <li>Changed medium for the cell line</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 18:</strong></p>
+<ul>
+  <li>Seeded HepG2 cells in 24-well plate for transfection of PEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP</li>
+  <li>Got polylysine for coating</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 19:</strong></p>
+<ul>
+  <li>Changed the medium for the HepG2 cell line</li>
+  <li>Seeded cells for another MTT assay</li>
+  <li>Checked HepG2 cells for transfection: growing slow, 30% confluency, cannot do transfection</li>
+  <li>Seeded cells on a 96-well plate for transfection in the coming week</li>
+  <li>Passaged HepG2 cells</li>
+</ul>
+<p>&nbsp;</p>
        </div>
     </div>
@@ Line 334: / Line 507: @@
     <div>
        <a href='#' class='head'>Week 4</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>July 22:</strong></p>
+<ul>
+  <li>Transfection of PEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP on 96-well plate</li>
+  <li>Added gradient concentration of sodium palmitate into HepG2 cells for MTT assay</li>
+  <li>Coated coverslips with polylysine for HepG2 cell staining and fixation</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 23:</strong></p>
+<ul>
+  <li>Replaced the medium after transfection of PEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP</li>
+  <li>Finished MTT assay to get cell viability in different FA concentrations in 24 hours</li>
+  <li>Transferred cells containing PEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP into 24-well plate with well-coated coverslips in each well</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 24:</strong></p>
+<ul>
+  <li>Stained mitochondria by Mito-tracker@Red and fixating by formaldehyde of HepG2 cells transfected with pEGFP-N1, pCMV/myc/mito/EGFP and <i>Pmv</i>/myc/mito/GFP in 24-well plate</li>
+  <li>Passaged HepG2 cells and changed medium of the cell line</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 25:</strong></p>
+<ul>
+  <li>Got results for cell staining and fixation: No obvious overlapping pattern between GPF signal and Mito-tracker red.</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 26:</strong></p>
+<ul>
+  <li>Coated coverslips by polylysine</li>
+  <li>Seeded HepG2 cells in one 24-well plate for transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP </li>
+  <li>Seeded HepG2 cells on two 96-well plate, for transfection of FABP construct and MTT assay in 48 hours</li>
+</ul>
+<p>&nbsp;</p>
        </div>
     </div>
@@ Line 340: / Line 546: @@
     <div>
        <a href='#' class='head'>Week 5</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>July 29:</strong></p>
+<ul>
+  <li>Transfection of FABP construct with pEGFP-N1 as control</li>
+  <li>Transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP in 24-well plate with well-coated coverslips</li>
+  <li>Added gradient concentration form 0.1mM to 1.2mM of FA into the plate for MTT assay</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>July 30:</strong></p>
+<ul>
+  <li>Changed medium after transfection of pEGFP-N1 and FABP construct and saw the results: Both pEGFP-N1 and FABP construct did not show GFP signals</li>
+  <li>Changed medium after transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP</li>
+  <li>Did staining and fixation of the coverslips and saw the results: No obvious overlapping patterns of GFP signals and Mito-tracker red</li>
+  <li>Passaged HepG2 cells</li>
+</ul>
+<p>&nbsp;</p>
        </div>
     </div>
@@ Line 349: / Line 571: @@
 <div>
        <a href='#' class='head'>Week 1</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>Aug 1:</strong></p>
+<ul>
+  <li>Seeded cells for transfection of pEGFP-N1, FABP construct, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP</li>
+  <li>Changed medium for HepG2 cell line</li>
+  <li>Got results for MTT assay in 48 hours</li>
+</ul><br>
+<p><strong>Aug 2:</strong></p>
+<ul>
+  <li>Checked if transfection (after 22 hrs) worked </li>
+  <ul>
+    <li>No GFP signal for pEFGP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP</li>
+  </ul>
+  <p>&nbsp;</p>
+</ul>
        </div>
      </div>
      <div>
        <a href='#' class='head'>Week 2</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>Aug 5: </strong></p>
+<ul>
+  <li>Transfection of HepG2 cells on 10cm Plates </li>
+  <li>Passaged P(n+8) HepG2 cells into one P(n+9)</li>
+  <li>Seeded HepG2 cells on 96-well plates for transfection </li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 6:</strong></p>
+<ul>
+  <li>Trouble shooting for  transfection of HepG2 cells on 96 well plates </li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 7: </strong></p>
+<ul>
+  <li>Observe transfected cell. GFP signal expressed </li>
+  <li>Seeded cells for transfection of pEGFP-N1, and (FABP construct/EGFP) in 96 well-plate </li>
+  <li>Seeded cells for transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP in 24 well plate with polylysine coated coverslips </li>
+  <li>Seeded cells for MTT assay </li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 8:</strong></p>
+<ul>
+  <li>Observed transfection of pEGFP-N1, and (FABP construct/EGFP) in 96 well-plate. No GFP signal observed</li>
+  <li>Observed transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP. GFP signal observed</li>
+  <li>Stained mitochondria of cells transfected with pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP using mito tracking dye </li>
+  <li>Observed cell with both transfection and stained mitochondria: No GFP signal observed because transfection occurred with very low efficiency. </li>
+  <li>Prepared new cell line: HEK293FT </li>
+  <li>Conducted MTT assay </li>
+</ul>
+<p>&nbsp;</p>
        </div>
      </div>
      <div>
        <a href='#' class='head'>Week 3</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>Aug 12: </strong></p>
+<ul>
+  <li>MTT Assay without seeding and incubation </li>
+  <li>Changed medium for HepG2 and HEK293FT cell lines </li>
+  <li>Prepared polylysine coated coverslips </li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 13:</strong></p>
+<ul>
+  <li>Seeded cells using HEK293FT cell line for transfection </li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 15:</strong></p>
+<ul>
+  <li>Cells detached and not in good condition for transfection </li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 16:</strong></p>
+<ul>
+  <li>Seeded cells HEK293FT cell line for transfection </li>
+</ul>
+<p>&nbsp;</p>
        </div>
      </div>
      <div>
        <a href='#' class='head'>Week 4</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>Aug 19:</strong></p>
+<ul>
+  <li>Transfection of pEGFP-N1, and (FABP construct/EGFP) in 96 well plate and pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP in 24 well plate.</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 20:</strong></p>
+<ul>
+  <li>Seeing results of transfection of FABP construct and pEGFP-N1 in HEK293FT cells and seeing the results: pEGFP-N1 had signal but FABP did not show GFP signals</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 22:</strong></p>
+<ul>
+  <li>Transfection of pEGFP-N1 and FABP construct into HEK293FT cells</li>
+  <li>Changing medium for HEK293FT cells and HepG2 cells and seeing the results: pEGFP-N1 had signal but FABP did not show GFP signals</li>
+</ul>
+<p>&nbsp;</p>
+      </div>
+    </div>
+  <div>
+      <a href='#' class='head'>Week 5</a>
+      <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>Aug 26:</strong></p>
+<ul>
+  <li>Transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP in 60mm TC plates</li>
+  <li>Changing medium of transfection of the previous day and seeing the results: pEGFP-N1 had signal but FABP did not show GFP signals</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 28:</strong></p>
+<ul>
+  <li>Changing medium of transfection of the previous day</li>
+  <li>Staining and fixation of </li>
+  <li>Drawing MTT standard curves</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 29:</strong></p>
+<ul>
+  <li>Transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP into HEK293FT cells</li>
+  <li>Maintaining HEK293FT and HepG2 cell line</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 30:</strong></p>
+<ul>
+  <li>Staining and fixation for HEK cells transfected by pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP</li>
+  <li>Seeing the results for transfected HEK293FT cells: No obvious overlapping pattern of GFP signals and Mito-tracker</li>
+  <li>Seeding HepG2 cells for Transfection of FABP construct and pEGFP-N1 in 96-well plate</li>
+  <li>Seeding HEK 293FT cells for transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP</li>
+  <li>Passaging HEK293FT cell line</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Aug 31:</strong></p>
+<ul>
+  <li>Transfection with pEGFP-N1 and pCMV/PolyA/EGFP into HEK293FT cells</li>
+</ul>
+<p>&nbsp;</p>
        </div>
      </div>
@@ Line 372: / Line 720: @@
      <div>
        <a href='#' class='head'>Week 1</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>Sept 1:</strong></p>
+<ul>
+  <li>Checking results for transfection on Aug 31: pEGFP-N1 had strong signal while PolyA did not have signals</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 2:</strong></p>
+<ul>
+  <li>Transfection of FABP construct and pEGFP-N1 in HepG2 cells</li>
+  <li>Check results for HEK293FT cells been transfected by pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP, by using confocal microscope</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 3:</strong></p>
+<ul>
+  <li>Adding FA into HepG2 cells been transfected by FABP construct and pEGFP-N1</li>
+  <li>Maintaining HepG2 and HEK293FT cell line</li>
+  <li>Drug selection Test on HEK293FT test by purmycin</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 4:</strong></p>
+<ul>
+  <li>Drug selection test on HEK293FT cells by neomycin and puromycin</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 5:</strong></p>
+<ul>
+  <li>Staining and fixation of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP and seeing he results by confocal microscope</li>
+  <li>Making mew DMEM</li>
+  <li>Drug selection test observation</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 6:</strong></p>
+<ul>
+  <li>Transfection of FAPB and pEGFP-N1 into HepG2 cells in 60 mm plates</li>
+  <li>Maintaining cell line for HEK293FT cells</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 7:</strong></p>
+<ul>
+  <li>Checking results for transfection on Sept 6: pEGFP-N1 showed weak GFP signal, no signals for FABP</li>
+  <li>Change medium for drug selection test</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 8:</strong></p>
+<ul>
+  <li>Maintaining cell line for HEK293FT cells and HepG2 cells</li>
+</ul>
+<p>&nbsp;</p>
        </div>
      </div>
      <div>
        <a href='#' class='head'>Week 2</a>
-       <div class='content' align="left">Content 1
+       <div class='content' align="left">
+<p>&nbsp;</p>
+<p><strong>Sept 9:</strong></p>
+<ul>
+  <li>Checking transfection for FABP construct again: Weak signals were shown from the debris of the cells</li>
+  <li>Transfection of <i>AceA</i> construct</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 11:</strong></p>
+<ul>
+  <li>Seeding HEK293FT cells into 35mm plates for transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP</li>
+  <li>Cell splitting of ACE</li>
+</ul>
+<p>&nbsp;</p>
+<p><strong>Sept 12:</strong></p>
+<ul>
+  <li>Transfection of pEGFP-N1, pCMV/myc/mito/EGFP and pCMV/myc/mito/GFP into HEK293FT cells</li>
+</ul>
+<p>&nbsp;</p>
        </div>
      </div>

Team:Hong Kong HKUST/notebook/mod1

From 2013.igem.org

Latest revision as of 23:13, 27 September 2013

FA Quantification and Cell Viability Module's Notebook

June 2013

July 2013

August 2013

September 2013