Team:LCG-UNAM-Mexico/Resources/Protocols/Ligation

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(New page: To ligate two biobricks into a base vector. Part 1: 2 µl Part 2: 2 µl Destination Plasmid: 2 µl 10X T4 Ligase Buffer: 2 µl T4 Ligase: 1 µl H2O: 11 µl Incubate at room tempera...)
 
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To ligate two biobricks into a base vector.
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{{Template:LCG_top_Netscape}}<!--Do not remove the first and last lines in this page!-->
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Part 1: 2 µl
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Part 2: 2 µl
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To ligate two biobricks into a base vector.
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Destination Plasmid: 2 µl
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<br><br>
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10X T4 Ligase Buffer: 2 µl
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Part 1: 2 µl<br>
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T4 Ligase: 1 µl
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Part 2: 2 µl<br>
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H2O: 11 µl
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Destination Plasmid: 2 µl<br>
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10X T4 Ligase Buffer: 2 µl<br>
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T4 Ligase: 1 µl<br>
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H2O: 11 µl<br>
Incubate at room temperature for 10 minutes and then inactivate the enzyme at 80°C for 20 minutes.
Incubate at room temperature for 10 minutes and then inactivate the enzyme at 80°C for 20 minutes.
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Transform 5 μl of the ligation product into 50 μl of competent DH5-alpha E. coli cells.
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[[Team:LCG-UNAM-Mexico/Resources/Protocols/Transformation|Transform]] 5 μl of the ligation product into 50 μl of competent DH5-alpha E. coli cells.
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<!--Do not remove the first and last lines in this page!-->{{Template:LCG_bottom_Netscape}}

Latest revision as of 19:11, 21 October 2009


To ligate two biobricks into a base vector.

Part 1: 2 µl
Part 2: 2 µl
Destination Plasmid: 2 µl
10X T4 Ligase Buffer: 2 µl
T4 Ligase: 1 µl
H2O: 11 µl

Incubate at room temperature for 10 minutes and then inactivate the enzyme at 80°C for 20 minutes.


Transform 5 μl of the ligation product into 50 μl of competent DH5-alpha E. coli cells.



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