Team:SDU-Denmark/labnotes2

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(Restriction digest of FlhDCmut and pSB1C3)
(Restriction digest of FlhDCmut and pSB1C3)
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''Notes:'' <br><br>
''Notes:'' <br><br>
''Results:''<br><br>
''Results:''<br><br>
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=== Restriction digest of FlhDCmut and pSB1C3 ===
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=== Restriction digest of FlhDCmut and pSB1C3 X2 ===
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''Experiment done by:'' Sheila <br><br>
''Experiment done by:'' Sheila <br><br>
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The digested samples were then run on a 1,5% agarose gel for approximately 20 minutes
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The digested samples were then run on a 1,5% agarose gel for approximately 20 minutes<br><br>
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''Results:'' The gel do not show any sign of a successful digest. Because the restiction enzymes only cut in the prefix and suffix of the FlhDCmut operon, it is not surprising that we do not see any bands of approximately 10bp length. However, the digestion should be very clearly visible in the plasmid backbone, as we should be cutting out RFP, which is approximately 1000bp long. The plasmid do not appear to have been digested by the restriction enzymes. 2 bands are visible, but they are both much longer than expected, in fact they appear around the length of the complete plasmid. So this experiment appears to have been unsuccessful.<br><br>
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''Results:'' Because it failed the first time, this experiment was repeated, with the same results (see notes section)<br><br>
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'''The first gel'''<br><br>
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'''The second gel'''<br><br>
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The gels do not show any sign of a successful digest. Because the restiction enzymes only cut in the prefix and suffix of the FlhDCmut operon, it is not surprising that we do not see any bands of approximately 10bp length. However, the digestion should be very clearly visible in the plasmid backbone, as we should be cutting out RFP, which is approximately 1000bp long. The plasmid do not appear to have been digested by the restriction enzymes. 2 bands are visible, but they are both much longer than expected, in fact they appear around the length of the complete plasmid. So this experiment appears to have been unsuccessful.<br><br>
''Notes:'' First troubleshooting session led to a repeat of the experiment, as we found that human error must have been the reason for the failed results, because it was the first time i performed the experiment. However, when the repeat of the restriction digest yielded the same results, we took a closer look and found that the combination of restriction enzymes was wrong. They were the combination we will use when we begin the assembly... In this case we want to insert the biobrick (FlhDCmut) into the plasmid backbone (pSB1C3, in which case we need to use the same restriction enzymes on both the plasmid and the biobrick. <br><br>
''Notes:'' First troubleshooting session led to a repeat of the experiment, as we found that human error must have been the reason for the failed results, because it was the first time i performed the experiment. However, when the repeat of the restriction digest yielded the same results, we took a closer look and found that the combination of restriction enzymes was wrong. They were the combination we will use when we begin the assembly... In this case we want to insert the biobrick (FlhDCmut) into the plasmid backbone (pSB1C3, in which case we need to use the same restriction enzymes on both the plasmid and the biobrick. <br><br>
--[[User:Sheila|Sheila]] 22:25, 1 August 2010 (UTC)
--[[User:Sheila|Sheila]] 22:25, 1 August 2010 (UTC)

Revision as of 14:14, 2 August 2010