G.tigrina Hox gene DthoxC insertion into prokaryote E.coli / (by UNIamCloning)/2011/10/06

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Fourth DNA Isolation (Gt.5-1 and Gt.5-2) and Second PCR (Gt.3 and Gt.4)

  • Today Alex and I started a fourth DNA isolation using two samples of planarians. Each sample contained approximately 0.012g of planarian. When the DNA extraction is performed, one sample will use the usual 30µl Buffer AE and the other will use 100µl Buffer AE to see if this affects the final DNA yield. Secondly, we set up and ran a second PCR using 2 samples of Gt. 3 and 3 samples of Gt. 4. For each sample, we increased the amount of DNA used from 1µl to 3µl and decreased the amount of water from 6.9µl to 5.4µl per sample. The annealing temperatures for the two Gt.3 samples were 52 and 51 degrees Celsius. The annealing temperatures for the three Gt.4 samples were 52, 51, and 50 degrees Celsius. For tomorrow, we will finish the DNA extraction for Gt.5-1 and Gt.5-2 and run it through the Qubit to check the DNA concentrations.