Difference between revisions of "Part:BBa K274004:Experience"
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== '''Characterisation by 2011 iGEM NCTU_FORMOSA''' == | == '''Characterisation by 2011 iGEM NCTU_FORMOSA''' == | ||
| − | + | we found out that there's point mutation in this plasmid. This mutation occurs in the Xba I restriction site which takes a lot of time for us just to digest it in order to ligase this part with our interest part. | |
| + | we then design a short sequence of primer. This primer include the correct sequence of the Xba I restriction site. We then do Polymerase Chain Reaction to extend the remaining sequence and amplify the correct plasmid. | ||
| + | The following figure shows the unsual gel electrophoresis result which the Xba I restriction site couldn't recognize the restriction site. | ||
We mutated the sequence to correct the Xba I restriction site . | We mutated the sequence to correct the Xba I restriction site . | ||
And the sequence is made of vioABDE not vioABCE | And the sequence is made of vioABDE not vioABCE | ||
Revision as of 03:49, 1 October 2011
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_K274004
User Reviews
UNIQdb448ddb51f14f6b-partinfo-00000000-QINU UNIQdb448ddb51f14f6b-partinfo-00000001-QINU
E.coli turned dark green after transformation of this plasmid, it is not supposed to be so since there is no promoter upstream of the operon. We have not found the reason.
Karina Arnesen, undergraduate, Alberta iGEM 2010
When digested with BamHI and NotI, the insert did not digest as expected, only a 2kb band (backbone) and ~6kb band were visible.
Characterisation by 2011 iGEM NCTU_FORMOSA
we found out that there's point mutation in this plasmid. This mutation occurs in the Xba I restriction site which takes a lot of time for us just to digest it in order to ligase this part with our interest part. we then design a short sequence of primer. This primer include the correct sequence of the Xba I restriction site. We then do Polymerase Chain Reaction to extend the remaining sequence and amplify the correct plasmid. The following figure shows the unsual gel electrophoresis result which the Xba I restriction site couldn't recognize the restriction site.
We mutated the sequence to correct the Xba I restriction site .
And the sequence is made of vioABDE not vioABCE