Difference between revisions of "Part:BBa J61001:Experience"
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The colonies were counted in each plate and the transformation efficiency was estimated in '''[CFU/ug of DNA]''' as: | The colonies were counted in each plate and the transformation efficiency was estimated in '''[CFU/ug of DNA]''' as: | ||
− | efficiency [CFU/ug of DNA]= # CFU * 1000 ng of DNA / amount of transformed DNA [ng] | + | <div align=center> |
+ | ''efficiency [CFU/ug of DNA]= # CFU * 1000 ng of DNA / amount of transformed DNA [ng]'' | ||
+ | </div> | ||
The results are shown here: | The results are shown here: |
Revision as of 17:24, 19 October 2010
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Applications of BBa_J61001
User Reviews
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••••
UNIPV-Pavia iGEM 2010 |
BBa_K300008 was used as a validation construct for this conditional replication origin, in order to test its capability to be propagated in pir+ or pir-116 strain and its inability to propagate in the other E. coli strains. In particular, BBa_K300008 was cut with XbaI-SpeI and the insert was isolated and purified from a 1% agarose gel. Then, it was self-ligated to generate a Cm-resistant R6K plasmid). BW25141 (BBa_K300984) and BW23474 (BBa_K300985) were chosen as pir+ and pir-116 strains respectively, while DH5alpha (BBa_V1001) and MC1061 (BBa_K300078) were chosen as pir- strains.
and plated on LB+Cm at 34 ug/ml.
efficiency [CFU/ug of DNA]= # CFU * 1000 ng of DNA / amount of transformed DNA [ng] The results are shown here:
These results show that BBa_J61001 replication origin can be only propagated in pir+ or pir-116 strains (BBa_K300084 and BBa_K300085), while the transformation of other strains with the R6K plasmid yielded no colonies after transformation. Moreover, these results show that the R6K plasmid in pir+ and pir-116 strains was transformed with the same efficiency as the pSB*** positive control plasmid, demonstrating that the R6K origin doesn't give any handicap in plasmid transformation. |
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