Difference between revisions of "Part:pSB3C5:Experience"
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<partinfo>pSB3C5</partinfo><br>
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<partinfo>pSB3C5 AddReview 4</partinfo><br>
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<I>Reshma Shetty</I>
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<I>[[User:Rshetty|Reshma Shetty]]</I>
 
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This plasmid seems to work.  When transformed into TOP10 cells, pSB3C5-I52001 does not transform but when cut with NotI and self-ligated (to eliminate ccdB), lots of colonies are obtained.  However, when miniprepped, pSB3C5-I52001 does not give as high of yields as expected for a high copy plasmid likely due to mutations in BBa_I52001 that render it nonfunctional as a high copy origin.  I am working to address this issue.  This plasmid should still work fine (it is just not as convenient to prep).
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[[Part:pSB3C5|pSB3C5]] is a functional low to medium copy plasmid but does not propagate at high copy.  When transformed into TOP10 cells, [[Part:pSB3C5|pSB3C5]]-[[Part:BBa_I52001|I52001]] does not transform as expected.  When [[Part:pSB3C5|pSB3C5]]-[[Part:BBa_I52001|I52001]] is cut with NotI,  self-ligated (to eliminate ''ccdB'' positive selection marker) and tranformed into TOP10, lots of colonies are obtained.  However, pSB3C5-I52001 does not give high miniprep yields due to mutations in BBa_I52001 that render it nonfunctional as a high copy origin.  [[Part:pSB3C5|pSB3C5]]-[[Part:BBa_I52001|I52001]] works fine as a plasmid but is not as convenient for plasmid DNA purification.
 
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Revision as of 01:00, 25 January 2008

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 pSB3C5

User Reviews

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••••

Reshma Shetty

pSB3C5 is a functional low to medium copy plasmid but does not propagate at high copy. When transformed into TOP10 cells, pSB3C5-I52001 does not transform as expected. When pSB3C5-I52001 is cut with NotI, self-ligated (to eliminate ccdB positive selection marker) and tranformed into TOP10, lots of colonies are obtained. However, pSB3C5-I52001 does not give high miniprep yields due to mutations in BBa_I52001 that render it nonfunctional as a high copy origin. pSB3C5-I52001 works fine as a plasmid but is not as convenient for plasmid DNA purification.

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