Today we prepared solutions, materials and the pre-inoculum to prepare electrocompetent E. coli.
Hemolysin Operon Amplification
We did the PCR to amplify the hemolysin operon using the total DNA extration that we had made before. Our sequence have almost 4000bp so we used a extension time of 3 minutes, the annelling temperatures in Celsius degrees was 56, 54, 52 and 51. No DNA fragment was shown in the agarose gel.
The terminator plasmid was quantified and we made another digestion, Protocol 14. We found by electrophoresis that it worked!