Difference between revisions of "Part:BBa K274004:Experience"

(Characterisation by 2011 iGEM NCTU_FORMOSA)
(Characterisation by 2011 iGEM NCTU_FORMOSA)
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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 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.
 
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 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.
 +
 
 +
 
  
 
And the sequence is made of vioABDE not vioABCE
 
And the sequence is made of vioABDE not vioABCE

Revision as of 03:50, 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

UNIQ854e7f1d344ec049-partinfo-00000000-QINU UNIQ854e7f1d344ec049-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. The following figure shows the unsual gel electrophoresis result which the Xba I restriction site couldn't recognize the restriction site.


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.


And the sequence is made of vioABDE not vioABCE