Team:Saarland/Test

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<h1>Methods</h1>
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<iframe src="//player.vimeo.com/video/109005558" width="500" height="281" frameborder="0" webkitallowfullscreen mozallowfullscreen allowfullscreen></iframe> <p><a href="http://vimeo.com/109005558">Saarland University iGEM 2014 - Campus Tour</a> </p></html>
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<h2>1. Cloning</h2>
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<h3>1.1 Restriction and phosphatase treatment of plasmid DNA</h3>
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Originally found in bacteria, endonucleases belong to a class of enzymes that have the ability to cleave the phosphodiester bond of DNA molecules by hydrolysis. Depending on the type they recognize site specific base sequences of 4, 6 or 8 bases that are often palindromic. Treatment of different DNA molecules with the same endonucleases allows the combination of the resulting DNA fragments in a following ligation reaction. <br><br>
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The treatment of DNA fragments, in particular plasmid DNA with phosphatase is beneficial if the restriction was performed with only one restriction enzyme or if the restriction with two different restriction enzymes was unbalanced and single cut plasmid DNA is left. As the nomenclature indicates the phosphatase cleaves the phosphate group at the 5´-terminus of DNA fragments and thereby minimizes the background of mono ligated plasmid DNA. Consequently the amount of transformants with insert DNA in the plasmid construct is considerably increased.<br><br>
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The following components were used for a restriction batch by default:<br>
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Latest revision as of 12:39, 17 October 2014


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