G.tigrina Hox gene DthoxC insertion into prokaryote E.coli / (by UNIamCloning)/2011/10/04

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PCR Check and Third DNA Isolation of G.tigrina

  • Today in lab we began by running a 1.0% agarose/TBE buffer electrophoresis gel to check success of last week's PCR amplification of our DthoxC planarian gene.

We then measured out approximately 0.025g of G.tigrina - (whole organism will readily lyse in proteinase K and ATL buffer) - because this quantity is suggested by our DNA extraction kit, Qiagen DNeasy Blood & Tissue Kit - (for 20ul proteinase K and 180ul ATL buffer). This involved weighing out about 15-20 planarians, which is considerably more than we used for any previous extractions (1-2 organisms). The 0.025g sample was scraped into a 1.5mL tube with 20ul proteinase K and 180ul ATL buffer, vortexed, and placed into a 56°C water bath overnight. The sample will continue to be isolated and purified tomorrow, Wednesday 10/05/2011.