Difference between revisions of "Part:BBa K2943902:Experience"

 
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<partinfo>BBa_K2943902</partinfo>
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<I>TAU_Israel</I>
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<b>Part Characterization:</b><br>
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This part is an improved version of an existing part (BBa_K608014). The mutations were detected by using bioinformatics softwares, and then the mutated part was ordered as gblock. Next, we proceeded to wet lab work. We started our lab work by transforming BBa_K608014 from the distribution kit to <em>E. coli</em> DH10beta. Then, after growing starters from the transformation plates overnight, we extracted the plasmids using miniprep.<br>
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Using PCR reaction, we amplified part of the plasmid, excluding part of the mRFP and RBS. This enabled us to insert the gblocks into the plasmid which included already the appropriate backbone and overlaps for the assembly.
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The gblock was inserted using Gibson Assembly. Before the assembly, we have done DPN1 to ensure no unwanted DNA template was present and used PCR clean kit to prepare the amplified backbone. Then we proceeded to the Gibson reaction and transformed the resulting plasmid into <em> E. coli</em> DH10beta.
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Revision as of 14:31, 20 October 2019


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Applications of BBa_K2943902

User Reviews

UNIQ8ab46bdd1a609b3b-partinfo-00000000-QINU UNIQ8ab46bdd1a609b3b-partinfo-00000001-QINU


BBa_K2943902 TAU_Israel

Part Characterization:
This part is an improved version of an existing part (BBa_K608014). The mutations were detected by using bioinformatics softwares, and then the mutated part was ordered as gblock. Next, we proceeded to wet lab work. We started our lab work by transforming BBa_K608014 from the distribution kit to E. coli DH10beta. Then, after growing starters from the transformation plates overnight, we extracted the plasmids using miniprep.

Using PCR reaction, we amplified part of the plasmid, excluding part of the mRFP and RBS. This enabled us to insert the gblocks into the plasmid which included already the appropriate backbone and overlaps for the assembly. The gblock was inserted using Gibson Assembly. Before the assembly, we have done DPN1 to ensure no unwanted DNA template was present and used PCR clean kit to prepare the amplified backbone. Then we proceeded to the Gibson reaction and transformed the resulting plasmid into E. coli DH10beta.

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