Team:WHU-China/Notebook/Protocols
From 2011.igem.org
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color:blue'>Preparation of competent cells (Using Calcium Chloride)</span></u></p></a> | color:blue'>Preparation of competent cells (Using Calcium Chloride)</span></u></p></a> | ||
+ | <a href="#upm"> | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
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font-family:黑体;color:blue'>(</span></u><u><span lang=EN-US style='font-size: | font-family:黑体;color:blue'>(</span></u><u><span lang=EN-US style='font-size: | ||
14.0pt;font-family:"Times New Roman","serif";color:blue'>Using Plasmid Mini Kit</span></u><u><span | 14.0pt;font-family:"Times New Roman","serif";color:blue'>Using Plasmid Mini Kit</span></u><u><span | ||
- | style='font-size:14.0pt;font-family:黑体;color:blue'>)</span></u></p> | + | style='font-size:14.0pt;font-family:黑体;color:blue'>)</span></u></p></a> |
+ | <a href="#tra"> | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
color:blue'>•<span style='font:7.0pt "Times New Roman"'> | color:blue'>•<span style='font:7.0pt "Times New Roman"'> | ||
</span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | </span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
- | color:blue'>Transformation (heat-shock method)</span></u></p> | + | color:blue'>Transformation (heat-shock method)</span></u></p></a> |
+ | <a href="#cwt"> | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
color:blue'>•<span style='font:7.0pt "Times New Roman"'> | color:blue'>•<span style='font:7.0pt "Times New Roman"'> | ||
</span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | </span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
- | color:blue'>Cleavage with two restriction enzymes</span></u></p> | + | color:blue'>Cleavage with two restriction enzymes</span></u></p></a> |
+ | <a href="#ge"> | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
color:blue'>•<span style='font:7.0pt "Times New Roman"'> | color:blue'>•<span style='font:7.0pt "Times New Roman"'> | ||
</span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | </span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
- | color:blue'> | + | color:blue'>Get Extraction</span></u></p></a> |
+ | <a href="#iod"> | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
color:blue'>•<span style='font:7.0pt "Times New Roman"'> | color:blue'>•<span style='font:7.0pt "Times New Roman"'> | ||
</span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | </span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
- | color:blue'>Identification of DNA by Electrophoresis</span></u></p> | + | color:blue'>Identification of DNA by Electrophoresis</span></u></p></a> |
+ | <a href="#li"> | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
color:blue'>•<span style='font:7.0pt "Times New Roman"'> | color:blue'>•<span style='font:7.0pt "Times New Roman"'> | ||
</span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | </span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
- | color:blue'>Ligation</span></u></p> | + | color:blue'>Ligation</span></u></p></a> |
+ | <a href="#pot" > | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
color:blue'>•<span style='font:7.0pt "Times New Roman"'> | color:blue'>•<span style='font:7.0pt "Times New Roman"'> | ||
</span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | </span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
- | color:blue'>PCR of the colony</span></u></p> | + | color:blue'>PCR of the colony</span></u></p></a> |
+ | <a href="#iofs"> | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
color:blue'>•<span style='font:7.0pt "Times New Roman"'> | color:blue'>•<span style='font:7.0pt "Times New Roman"'> | ||
</span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | </span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
- | color:blue'>Identification of DNA by Sequencing</span></u></p> | + | color:blue'>Identification of DNA by Sequencing</span></u></p></a> |
- | + | ||
+ | <a href="#ot"> | ||
<p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | <p class=MsoListParagraph style='margin-left:21.0pt;text-indent:-21.0pt'><span | ||
lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
color:blue'>•<span style='font:7.0pt "Times New Roman"'> | color:blue'>•<span style='font:7.0pt "Times New Roman"'> | ||
</span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | </span></span><u><span lang=EN-US style='font-size:14.0pt;font-family:"Times New Roman","serif"; | ||
- | color:blue'>Others</span></u></p> | + | color:blue'>Others</span></u></p></a> |
</div> | </div> | ||
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"Times New Roman","serif"'> </span></p> | "Times New Roman","serif"'> </span></p> | ||
+ | <a name="upm"> | ||
<p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | <p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | ||
style='font-size:20.0pt;font-family:宋体'>Ⅲ</span><span lang=EN-US | style='font-size:20.0pt;font-family:宋体'>Ⅲ</span><span lang=EN-US | ||
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Extraction</span><span style='font-size:20.0pt;font-family:黑体'>(</span><span | Extraction</span><span style='font-size:20.0pt;font-family:黑体'>(</span><span | ||
lang=EN-US style='font-size:20.0pt;font-family:"Times New Roman","serif"'>Using | lang=EN-US style='font-size:20.0pt;font-family:"Times New Roman","serif"'>Using | ||
- | Plasmid Mini Kit</span><span style='font-size:20.0pt;font-family:黑体'>)</span></p> | + | Plasmid Mini Kit</span><span style='font-size:20.0pt;font-family:黑体'>)</span></p></a> |
<p class=MsoNormal><span lang=EN-US style='font-size:15.0pt;font-family:"Times New Roman","serif"'>3.1 | <p class=MsoNormal><span lang=EN-US style='font-size:15.0pt;font-family:"Times New Roman","serif"'>3.1 | ||
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280nm.</span></p> | 280nm.</span></p> | ||
+ | <a name="tra"> | ||
<p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | <p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | ||
style='font-size:20.0pt;font-family:宋体'>Ⅳ</span><span lang=EN-US | style='font-size:20.0pt;font-family:宋体'>Ⅳ</span><span lang=EN-US | ||
style='font-size:20.0pt;font-family:"Times New Roman","serif"'>. Transformation | style='font-size:20.0pt;font-family:"Times New Roman","serif"'>. Transformation | ||
- | (heat-shock method)</span></p> | + | (heat-shock method)</span></p></a> |
<p class=MsoListParagraph style='margin-left:21.3pt;text-indent:-21.3pt; | <p class=MsoListParagraph style='margin-left:21.3pt;text-indent:-21.3pt; | ||
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at 37</span><span style='font-size:12.0pt;line-height:150%;font-family:宋体'>℃</span></p> | at 37</span><span style='font-size:12.0pt;line-height:150%;font-family:宋体'>℃</span></p> | ||
+ | <a name="cwt"> | ||
<p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | <p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | ||
style='font-size:20.0pt;font-family:宋体'>Ⅴ<span lang=EN-US>.Cleavage with two | style='font-size:20.0pt;font-family:宋体'>Ⅴ<span lang=EN-US>.Cleavage with two | ||
- | restriction enzymes</span></span></p> | + | restriction enzymes</span></span></p></a> |
<p class=MsoListParagraph style='text-indent:2.0cm'><b><span lang=EN-US | <p class=MsoListParagraph style='text-indent:2.0cm'><b><span lang=EN-US | ||
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font-family:"Times New Roman","serif"'>Quick operation is recommended.</span></p> | font-family:"Times New Roman","serif"'>Quick operation is recommended.</span></p> | ||
+ | <a name="ge"> | ||
<p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | <p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | ||
style='font-size:20.0pt;font-family:宋体'>Ⅵ</span><span lang=EN-US | style='font-size:20.0pt;font-family:宋体'>Ⅵ</span><span lang=EN-US | ||
- | style='font-size:20.0pt;font-family:"Times New Roman","serif"'>. | + | style='font-size:20.0pt;font-family:"Times New Roman","serif"'>. Get Extraction</span></p></a> |
<p class=MsoNormal style='line-height:150%'><b><span lang=EN-US | <p class=MsoNormal style='line-height:150%'><b><span lang=EN-US | ||
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280nm.</span></p> | 280nm.</span></p> | ||
+ | <a name="iod"> | ||
<p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | <p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | ||
style='font-size:20.0pt;font-family:宋体'>Ⅶ</span><span lang=EN-US | style='font-size:20.0pt;font-family:宋体'>Ⅶ</span><span lang=EN-US | ||
style='font-size:20.0pt;font-family:"Times New Roman","serif"'>. Identification | style='font-size:20.0pt;font-family:"Times New Roman","serif"'>. Identification | ||
- | of DNA by Electrophoresis</span></p> | + | of DNA by Electrophoresis</span></p></a> |
<p class=MsoListParagraph style='text-indent:24.1pt;line-height:150%'><b><span | <p class=MsoListParagraph style='text-indent:24.1pt;line-height:150%'><b><span | ||
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<b>DL 2000</b></span></p> | <b>DL 2000</b></span></p> | ||
+ | <a name="li"> | ||
<p class=MsoNormal align=left style='text-align:left'><span style='font-size: | <p class=MsoNormal align=left style='text-align:left'><span style='font-size: | ||
20.0pt;font-family:宋体'>Ⅷ</span><span lang=EN-US style='font-size:20.0pt; | 20.0pt;font-family:宋体'>Ⅷ</span><span lang=EN-US style='font-size:20.0pt; | ||
- | font-family:"Times New Roman","serif"'>. Ligation</span></p> | + | font-family:"Times New Roman","serif"'>. Ligation</span></p></a> |
<p class=MsoListParagraph style='text-indent:2.0cm'><b><span lang=EN-US | <p class=MsoListParagraph style='text-indent:2.0cm'><b><span lang=EN-US | ||
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style='font-family:"Times New Roman","serif"'> </span></p> | style='font-family:"Times New Roman","serif"'> </span></p> | ||
+ | <a name="pot"> | ||
<p class=MsoNormal align=left style='text-align:left'><span style='font-size: | <p class=MsoNormal align=left style='text-align:left'><span style='font-size: | ||
20.0pt;font-family:宋体'>Ⅸ</span><span lang=EN-US style='font-size:20.0pt; | 20.0pt;font-family:宋体'>Ⅸ</span><span lang=EN-US style='font-size:20.0pt; | ||
- | font-family:"Times New Roman","serif"'>. PCR of the colony</span></p> | + | font-family:"Times New Roman","serif"'>. PCR of the colony</span></p></a> |
<p class=MsoListParagraph style='text-indent:129.75pt'><b><span lang=EN-US | <p class=MsoListParagraph style='text-indent:129.75pt'><b><span lang=EN-US | ||
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PCR mix should be blended.</span></p> | PCR mix should be blended.</span></p> | ||
+ | <a name="iods"> | ||
<p class=MsoNormal><span style='font-size:20.0pt;font-family:宋体'>Ⅹ</span><span | <p class=MsoNormal><span style='font-size:20.0pt;font-family:宋体'>Ⅹ</span><span | ||
lang=EN-US style='font-size:20.0pt;font-family:"Times New Roman","serif"'>.Identification | lang=EN-US style='font-size:20.0pt;font-family:"Times New Roman","serif"'>.Identification | ||
- | of DNA by Sequencing</span></p> | + | of DNA by Sequencing</span></p></a> |
<p class=MsoNormal style='text-indent:42.0pt'><span lang=EN-US | <p class=MsoNormal style='text-indent:42.0pt'><span lang=EN-US | ||
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sequencing.</span></p> | sequencing.</span></p> | ||
+ | <a name="ot"> | ||
<p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | <p class=MsoNormal align=left style='margin-left:-.2pt;text-align:left'><span | ||
- | lang=EN-US style='font-size:20.0pt;font-family:"Times New Roman","serif"'>XI.Others</span></p> | + | lang=EN-US style='font-size:20.0pt;font-family:"Times New Roman","serif"'>XI.Others</span></p></a> |
<p class=MsoNormal align=left style='margin-right:18.0pt;text-align:left; | <p class=MsoNormal align=left style='margin-right:18.0pt;text-align:left; |
Revision as of 03:06, 23 September 2011