Part:BBa_M30109:Experience

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.

User Reviews

The Edinburgh 2010 team attempted to transform this biobrick repeatedly from the 2010 distribution. When this failed, we tried transforming the individual components (I15008, I15009, I15010). This also didn't work. We eventually got a result when we tried amplifying each individual part out of the full biobrick using combinations of 3 different sets of primers, one for each of the components. The only one which gave a PCR product was I15010 (the cph8), leading us to suspect that the M30109 part contains only the cph8 component and not the other two parts (ho1 and PcyA). We have been made aware that Mexico UNAM-Genomics 2010 are also struggling with this part, and we know of at least one other team who have plans to utilise it. We assume that they are also unsuccesful with this, but we request that they but their own experience up here if we are wrong.

If we can find a way to put the full thing back together, we will. If anyone else feels like attempting this, that would also be appreciated. Otherwise we highly recommend that you do not attempt to use this part until the problems with it have been resolved, and we suggest that the registry remove this particular biobrick from the general distribution, so that teams do not end up wasting time, energy and resources trying to get it to work.

The Caltech 2010 team also struggled to use this part. We eventually determined that the sequence of BBa_M30109 that we received from the Registry was completely wrong. Upon trying to sequence its constituent parts, we determined that the sequence of BBa_I15010 was incorrect, and may be (at least one) source of error.

The 2011 University of Minnesota iGEM team failed to transform this part into E. coli after two attempts. Known viable competent cells were used as these same cells were used to concurrently transform other registry parts. PCR amplification of this construct also failed.

The 2012 ETH Zurich team could transform it. However, the sequencing-result of this part shows a random sequence that is in no way related to the provided sequence. As the Edinburgh team we suggest to remove this part completely.

The 2012 SEU_O_China team could also transform it.However, BBa_M30109 seems not available，as the comments above.

UNIQbc78b18d314d88ca-partinfo-00000002-QINU UNIQbc78b18d314d88ca-partinfo-00000003-QINU

2014_WashU_StLouis

0 Stars out of 5! Sequence is NOT as expected. We attempted to use this part in order to activate the CcaR/CcaS light sensor system. Primers had mismatches. Band lengths for the biobrick part is as expected but the sequence is mostly garbage. We tried cloning this into two separate plasmids. We discarded both of them. Colony PCR showed wrong bands, Digestion of the biobrick part also failed. We ended up discarding this biobrick part in favor of another we found from the registry. Visit our project here: http://2014.igem.org/Team:WashU_StLouis

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