"
Team:Leicester/Parts
From 2012.igem.org
| Line 115: | Line 115: | ||
</br>^ all sequenced <i>Pseudomonas putida</i> strains had the same gene sequence | </br>^ all sequenced <i>Pseudomonas putida</i> strains had the same gene sequence | ||
</br> | </br> | ||
| - | </br> | + | </br> <div align="center"><img src="http:// 2012.igem.org/wiki/images/c/c3/Tod_B_large_gel_17.9.12.jpeg " /></div> |
</br> | </br> | ||
</br> TodB - 324bp | </br> TodB - 324bp | ||
Revision as of 13:57, 26 September 2012
Parts Submitted to the Registry
An important aspect of the iGEM competition is the use and creation of standard biological parts. Each team will make new parts during iGEM and will place them in the Registry of Standard Biological Parts. The iGEM software provides an easy way to present the parts your team has created . The "groupparts" tag will generate a table with all of the parts that your team adds to your team sandbox. Note that if you want to document a part you need to document it on the Registry, not on your team wiki.
Remember that the goal of proper part documentation is to describe and define a part such that it can be used without a need to refer to the primary literature. The next iGEM team should be able to read your documentation and be able to use the part successfully. Also, you should provide proper references to acknowledge previous authors and to provide for users who wish to know more.
Primers and PCR
Having worked out how long screening a DNA library could take we realised that it would be quicker to try to isolate our genes of interest using PCR. We used BLAST on sequenced Pseudomonas genomes to locate the genes of the Tod operon, and found the whole operon was remarkably conserved. Only a couple of the genes varied at all between the sequenced strains: we took account of this by adding a few degenerate positions in the PCR primers (see below). We concentrated on genes in the Tod operon that didn't have restriction sites incompatible with the basic Biobrick standard:
TodX: (544bp) >TODXF 5'-atgcccgccagtctgacgcttg-3' >TODXR 5'-accagccagcaccatgcggc-3' TodX gene in all putida the same regardless of strain
Unfortunately, with the 2 strains of Pseudomonas we had in the lab, none of the primers amplified a product of a size consistent with the target genes, meaning that the strains likely do not encode any of the Tod operon genes. As a result we couldn't make a biobrick, although the strategy should . However, these analyses and primers will be available for future iGEM Leicester teams. With the right strains of bacteria, or new bacteria isolated in the "Citizen Science Experiment", they may be able to extract the target genes relatively quickly, for biobrick construction and characterisation.
