Team:Cambridge/Experiments
From 2011.igem.org
Contents |
Experiments
Details of the experiments carried out throughout the project are linked from this page. These experiments should also be linked to from the appropriate diary page.
Preliminary Exercise
Initial exercise during our 2 weeks crash course in synthetic biology with the aim of familiarising us with common laboratory methods of preparing and assembling DNA.
Main Project - 'Bactiridescence'
Amplification of Reflectin Genes from the Squid Genomic DNA - Part 1
Reflectin genes were amplified directly from [http://en.wikipedia.org/wiki/Loligo Loligo] tissue. Tissue from the Loligo genus was commercially available from fishing bait suppliers and culinary wholesalers. Primers were designed from the nucleotide sequences of three reflectin proteins identified in L. pealei, and used in a PCR reaction upon L. vulgaris genomic DNA.
Amplification of Reflectin Genes from the Squid Genomic DNA - Part 2
Two new protocols for genomic DNA extraction were used in order to improve yield and purity of DNA. In addition to three sets of primers allowing for amplification of reflectin, an extra 'positive control' pair of primers was used in the PCR reaction.
Transformation of E.coli with Optimised Reflectin Plasmids
After failing to isolate reflectin genes from squid genomic DNA, we contacted several researchers who authored the papers we found in the literature about reflectin. Dr. Wendy Crookes-Goodson very kindly offered to donate a sample of codon-optimised reflectin genes that she used in her research. These arrived on cloning (non-expressing) plasmids The first part of this simple procedure involves extraction the plasmid DNA sent by Dr. Wendy Crookes-Goodson, a co-author of several important papers studying reflectin. The second part is transformation of E.coli with the plasmids DNA to store and amplify the plasmids DNA.