Difference between revisions of "Team:CPU CHINA/Experiments"
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<h2>Characterization</h2> | <h2>Characterization</h2> | ||
| − | <h4>You can click on these arrows to read the | + | <h4>You can click on these arrows to read the protocol .</h4> |
<center> | <center> | ||
<div> | <div> | ||
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</center> | </center> | ||
| − | <center><h4 style="font-size:unset !important">You can <a href="javascript:void(0);" onclick="showExperiment(5)">click here to see | + | <center><h4 style="font-size:unset !important">You can <a href="javascript:void(0);" onclick="showExperiment(5)">click here to see the protocol of promoter identification</a>.</h4></center> |
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"<h2>qPCR</h2>"+ | "<h2>qPCR</h2>"+ | ||
| − | "<h4>Follow the instruction in <i>Vazyme ChamQ Universal SYBR® qPCR Master Mix</i> (Vazyme Biotech, China).</h4><br>"; | + | "<h4>Follow the instruction in <a href=\"https://static.igem.org/mediawiki/2018/5/58/T--CPU_CHINA--experiment_qpcr2.pdf\"><i>Vazyme ChamQ Universal SYBR® qPCR Master Mix</i></a> (Vazyme Biotech, China).</h4><br>"; |
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var qidongzi = "<h2>Cell transfection and luciferase reporter gene assay</h2>"+ | var qidongzi = "<h2>Cell transfection and luciferase reporter gene assay</h2>"+ | ||
"<h4>pGL3-Basic was used as negative Control, and pGL3-Control was used as positive Control.</h2>"+ | "<h4>pGL3-Basic was used as negative Control, and pGL3-Control was used as positive Control.</h2>"+ | ||
| − | "<h4>HepG2 cells were transfected with 1ug of pGL3-htert, pGL3-hulc and pGL3-control. HepG2 cells were cultured in DEME medium containing 10% heat inactivated bovine serum at 37 ℃ and 5% CO2 constant temperature incubator. HepG2 cells were seeded in 96-well plate according to 2* 10<sup>5</sup>/ well, and cultured in non-antibiotic culture medium for 24 hours, then transfected with liposome (Lipo fectamin3000), and transfected with 3 holes per plasmid. The transfection medium was replaced after 4 to 6 h, and the DEME culture medium containing 10% calf serum was used to continue the culture. After 48 h, the cells were treated with Promege's <a><u>steady-glo Luciferase Assay System kit </u></a>and the activity of <i>Luc</i> was determined.</h4><br>"; | + | "<h4>HepG2 cells were transfected with 1ug of pGL3-htert, pGL3-hulc and pGL3-control. HepG2 cells were cultured in DEME medium containing 10% heat inactivated bovine serum at 37 ℃ and 5% CO2 constant temperature incubator. HepG2 cells were seeded in 96-well plate according to 2* 10<sup>5</sup>/ well, and cultured in non-antibiotic culture medium for 24 hours, then transfected with liposome (Lipo fectamin3000), and transfected with 3 holes per plasmid. The transfection medium was replaced after 4 to 6 h, and the DEME culture medium containing 10% calf serum was used to continue the culture. After 48 h, the cells were treated with Promege's <a href=\"https://static.igem.org/mediawiki/2018/9/9a/T--CPU_CHINA--experiment_promoter.pdf\"><u>steady-glo Luciferase Assay System kit </u></a>and the activity of <i>Luc</i> was determined.</h4><br>"; |
var zhuanran = "<h2>Cell transfection and Screening</h2>"+ | var zhuanran = "<h2>Cell transfection and Screening</h2>"+ | ||
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"<h2>Luciferase reporter gene assay</h2>"+ | "<h2>Luciferase reporter gene assay</h2>"+ | ||
| − | "<h4>After 48 h, follow the instruction in <a><i>steady-glo Luciferase Assay System kit</i></a> (Promege, US) to measure the activity of luciferase expressed from the plasmids. </h4><br>"; | + | "<h4>After 48 h, follow the instruction in <a href=\"https://static.igem.org/mediawiki/2018/9/9a/T--CPU_CHINA--experiment_promoter.pdf\"><i>steady-glo Luciferase Assay System kit</i></a> (Promege, US) to measure the activity of luciferase expressed from the plasmids. </h4><br>"; |
Latest revision as of 15:09, 7 December 2018
Experiments
Molecular Cloning
Transfection
You can click on the figure to see the protocol.
Characterization
You can click on these arrows to read the protocol .
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