Team:Bio Without Borders/Experiments

Experiments

We improved the empty cassette of TecCEM 2017 team to include BamHI and HincII sites and spacer DNA for the siRNA. We worked on cloning the empty cassette, BBa_K3277000 with a linearized chloramphenicol backbone from the iGEM Distribution kit, and an ampicillin backbone from a J04450-pSB1A3 plasmid. The empty cassette is a vector for our siRNAs targeting the chitin synthase and superoxide dismutase genes. Then, we ligated the BBa_3277000 + pSB1C3 plasmid with the oligos of the siRNAs. We also attempted to improve upon the WNT siRNA of the previous team.

Digestion of Linearized Backbones · Benchling
Ligation with T4 DNA Ligase · Benchling
Restriction Digest using EcoR1 and Pst1 · Benchling
Restriction Digest with BamHI and HincII · Benchling
Colony PCR (cPCR) with OneTaq 2X Mastermix · Benchling
PCR Protocol · Benchling
Annealing Oligos · Benchling