Team:ESBS-Strasbourg/Results/Assembly
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<div class="heading">A new standard for multiple Biobrick assembly</div> | <div class="heading">A new standard for multiple Biobrick assembly</div> | ||
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The ligation step creates several different products. This occurs because the NgoMIV and AgeI enzymes are restriction enzymes which cut the sequence as palindromes. This allows same fragments to be ligated together (number 4 and 5) which occurs especially with the short fragments (Number 1 and 3). Also the ligation of the short fragments cut with AgeI and NgoMIV can be ligated (number 2). All possible side products are shown in figure 2. | The ligation step creates several different products. This occurs because the NgoMIV and AgeI enzymes are restriction enzymes which cut the sequence as palindromes. This allows same fragments to be ligated together (number 4 and 5) which occurs especially with the short fragments (Number 1 and 3). Also the ligation of the short fragments cut with AgeI and NgoMIV can be ligated (number 2). All possible side products are shown in figure 2. | ||
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To avoid the amplification of side products after the ligation and to skip the separation step on the LMP gel, a nested PCR can be used. For this reaction others primers have to be designed and the primers VF and VR cannot be used. This different step can be used if two biobricks of the same size have to be fused together. | To avoid the amplification of side products after the ligation and to skip the separation step on the LMP gel, a nested PCR can be used. For this reaction others primers have to be designed and the primers VF and VR cannot be used. This different step can be used if two biobricks of the same size have to be fused together. |
Revision as of 23:24, 23 October 2010
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