Difference between revisions of "Part:BBa K1354002:Experience"

 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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<partinfo>BBa_K1354002 AddReview 4</partinfo>
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<I>Cooper Union iGEM 2014</I>
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Enter the review inofrmation here.
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[[File:CU_1017_TdT.png]]
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As seen from the polyacrylamide gel shown above, the TdT enzyme expressed in the Rosetta cells using our genetically engineered plasmid works as intended; base pairs are appended to oligos by the enzyme. However, there is a difference between the activity of our enzyme compared to the commercially obtained TdT. The majority of the oligos have only a few base pairs added by our expressed TdT while the lane of the commercial TdT is a smear of oligos, suggesting the commercial TdT has a greater reaction rate. But it can also be attributed to the fact that the 0.5 U/μL concentration of our expressed TdT is an overestimation. The TdT concentration of the reaction with our expressed TdT is less than that of the commercial TdT. <br><br>
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There is also a large gap between the oligos with only a few bases added and those with 30+ bases added. This suggests that our expressed TdT has a higher processivity than that of commercially obtained TdT. This might be due to subtle sequence differences such as the His tag on our TdT sequence.
 
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Revision as of 20:30, 1 November 2014

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1354002

User Reviews

UNIQ799b1a630784b40b-partinfo-00000000-QINU


••••

Cooper Union iGEM 2014

Enter the review inofrmation here.

;

CU 1017 TdT.png

As seen from the polyacrylamide gel shown above, the TdT enzyme expressed in the Rosetta cells using our genetically engineered plasmid works as intended; base pairs are appended to oligos by the enzyme. However, there is a difference between the activity of our enzyme compared to the commercially obtained TdT. The majority of the oligos have only a few base pairs added by our expressed TdT while the lane of the commercial TdT is a smear of oligos, suggesting the commercial TdT has a greater reaction rate. But it can also be attributed to the fact that the 0.5 U/μL concentration of our expressed TdT is an overestimation. The TdT concentration of the reaction with our expressed TdT is less than that of the commercial TdT.

There is also a large gap between the oligos with only a few bases added and those with 30+ bases added. This suggests that our expressed TdT has a higher processivity than that of commercially obtained TdT. This might be due to subtle sequence differences such as the His tag on our TdT sequence. UNIQ799b1a630784b40b-partinfo-00000002-QINU