Difference between revisions of "Part:BBa K4205112:Experience"
 
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<p>For our reporter gene lacZ, in order to determine whether our chassis bacteria ''Escherichia coli'' DH5α themselves carry this gene, which may interfere with the results of our functional analysis. We designed primers, performed colony PCR to amplify the lacZ gene, and ran agarose gel.</p>
 
<p>For our reporter gene lacZ, in order to determine whether our chassis bacteria ''Escherichia coli'' DH5α themselves carry this gene, which may interfere with the results of our functional analysis. We designed primers, performed colony PCR to amplify the lacZ gene, and ran agarose gel.</p>
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<p>As shown in the figure below, after colony PCR, the sample from the transformed DH5α ran out bands near 3000 bp, which was consistent with the target band (3075bp). The sample from the transformed DH5α can also run out bands near 3000 bp but with much brighter light. This indicated that our chassis bacteria Escherichia coli DH5α themselves carried the lacZ gene and could express them in micro amounts.</p>
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[[File:4-laczpcr.png|500px|center|thumb|left|PCR amplification of lacZ reporter gene.  
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[[File:4-laczpcr.png|500px|thumb|left|PCR amplification of lacZ reporter gene.  
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Lanes 1-2: PCR sample form transformed DH5α. Lanes 3-4: PCR sample form untransformed DH5α.
 
Lanes 1-2: PCR sample form transformed DH5α. Lanes 3-4: PCR sample form untransformed DH5α.
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<p>As shown in the figure above, after colony PCR, the sample from the transformed DH5α ran out bands near 3000 bp, which was consistent with the target band (3075bp). The sample from the transformed DH5α can also run out bands near 3000 bp but with much brighter light. This indicated that our chassis bacteria Escherichia coli DH5α themselves carried the lacZ gene and could express them in micro amounts.</p>

Latest revision as of 11:15, 13 October 2022


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For our reporter gene lacZ, in order to determine whether our chassis bacteria Escherichia coli DH5α themselves carry this gene, which may interfere with the results of our functional analysis. We designed primers, performed colony PCR to amplify the lacZ gene, and ran agarose gel.

PCR amplification of lacZ reporter gene. Lanes 1-2: PCR sample form transformed DH5α. Lanes 3-4: PCR sample form untransformed DH5α.

As shown in the figure above, after colony PCR, the sample from the transformed DH5α ran out bands near 3000 bp, which was consistent with the target band (3075bp). The sample from the transformed DH5α can also run out bands near 3000 bp but with much brighter light. This indicated that our chassis bacteria Escherichia coli DH5α themselves carried the lacZ gene and could express them in micro amounts.