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Team:Hong Kong HKUST/notebook/mod1
From 2013.igem.org
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li { position:relative;overflow:hidden; } | li { position:relative;overflow:hidden; } | ||
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| + | #hkust_Logo{ | ||
| + | width:60px; | ||
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</style> | </style> | ||
<style type="text/css"> | <style type="text/css"> | ||
| - | + | body{background-color:#494042;width:100%;margin-bottom:20px;min-width:600px;max-width:2000px;height:100%;font: x-small sans-serif;} | |
#menubar | #menubar | ||
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#p-logo | #p-logo | ||
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{display:none} | {display:none} | ||
#kepala | #kepala | ||
| - | {position:absolute;top:15px;background-color:#F0A5BE;height: | + | {position:absolute;top:15px;background-color:#F0A5BE;} |
| - | + | #cover | |
| - | { | + | {position:absolute;top:0px;height:168px;width:100%;background:#494042;z-index:-1;} |
| + | body, div, dl, dt, dd, ul, ol, li, h1, h2, h3, h4, h5, h6, pre, form, fieldset, p, blockquote, th, td { | ||
| + | margin: 0; | ||
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| + | div.menu | ||
| + | { | ||
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| + | width: 940px; | ||
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| + | |||
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| + | display: table; | ||
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.menu ul,li | .menu ul,li | ||
{ | { | ||
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list-style:none; | list-style:none; | ||
width:100%; | width:100%; | ||
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} | } | ||
ul.first>li | ul.first>li | ||
{ | { | ||
float:left; | float:left; | ||
| - | width:16. | + | width:16.6667%; |
| + | z-index:11; | ||
} | } | ||
ul.first>li>a:link,ul.first>li>a:visited | ul.first>li>a:link,ul.first>li>a:visited | ||
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text-decoration:none; | text-decoration:none; | ||
text-transform:uppercase; | text-transform:uppercase; | ||
| + | z-index:11; | ||
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ul.first>li>a:hover,ul.first>li>a:active | ul.first>li>a:hover,ul.first>li>a:active | ||
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{ | { | ||
display:none; | display:none; | ||
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text-decoration:none; | text-decoration:none; | ||
text-transform:uppercase; | text-transform:uppercase; | ||
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} | } | ||
ul.first>li>ul>li>a:hover,ul.first>li>ul>li>a:active | ul.first>li>ul>li>a:hover,ul.first>li>ul>li>a:active | ||
{ | { | ||
| - | background-color:#f87790; | + | background-color:#f87790;z-index:11; |
} | } | ||
.menu | .menu | ||
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top:141px; | top:141px; | ||
width:100%; | width:100%; | ||
| + | z-index:11; | ||
| + | } | ||
| + | ul.side-nav { | ||
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| + | padding: 17px 0; | ||
| + | } | ||
| + | ul.side-nav h6 { | ||
| + | margin: 47px 0px 0px; | ||
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| + | ul.side-nav li.divider { | ||
| + | border-top: 1px solid #ddd; | ||
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| + | .two, .row .two { | ||
| + | width: 16.66667%; | ||
| + | top:160px; | ||
} | } | ||
ul, ol { | ul, ol { | ||
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font-size: 14px; | font-size: 14px; | ||
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| + | } | ||
| + | [class*="column"] + [class*="column"]:last-child { | ||
| + | float: right; | ||
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| + | .ten, .row .ten { | ||
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| + | top:192px; | ||
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| + | } | ||
| + | .column, .columns { | ||
| + | float: left; | ||
| + | min-height: 1px; | ||
| + | padding: 0 15px; | ||
| + | position: relative; | ||
| + | } | ||
| + | .twelve, .row .twelve { | ||
| + | width: 100%; | ||
| + | } | ||
| + | h1, h2, h3, h4, h5, h6 { | ||
| + | font-family:"Trebuchet MS", Helvetica, sans-serif | ||
| + | font-weight: normal; | ||
| + | color: #000; | ||
| + | border-bottom: none; | ||
| + | } | ||
| + | div.team-bios-container div.row { | ||
| + | padding: 0px 0px 10px; | ||
| + | border-bottom: 1px solid #ddd; | ||
| + | } | ||
| + | .row .row { | ||
| + | width: auto; | ||
| + | max-width: none; | ||
| + | min-width: 0; | ||
| + | margin: 0 -15px; | ||
| + | } | ||
| + | media="screen, projection" | ||
| + | #bodyContent h1, #bodyContent h2 { | ||
| + | margin-bottom: .6em; | ||
| + | } | ||
| + | h1.centered, h2.centered, h3.centered, h4.centered, h5.centered, h6.centered { | ||
| + | text-align: center; | ||
| + | } | ||
| + | h2 { | ||
| + | font-size: 37px; | ||
| + | } | ||
| + | .nine, .row .nine { | ||
| + | width: 75%; | ||
| + | } | ||
| + | [class*="column"] + [class*="column"]:last-child { | ||
| + | float: right; | ||
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| + | .three, .row .three { | ||
| + | width: 25%; | ||
} | } | ||
#flight{z-index:-2;} | #flight{z-index:-2;} | ||
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<body> | <body> | ||
| + | <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"> | <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">Notebook</a></li> | ||
| - | <li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/notebook/ | + | <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/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/mod3">Module 3</a></li> | ||
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<li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/Wetlab">Wetlab</a> | <li><a href="https://2013.igem.org/Team:Hong_Kong_HKUST/Wetlab">Wetlab</a> | ||
<ul> | <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> | ||
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
