PCR of GFP and ATF1 plasmids
- Today we diluted the primers we ordered and set up a PCR using them and the Vf and Vr primers taken from our Biobrick parts. We used Vf and 15_E0040_R to amplify the GFP gene and for the ATF1 gene we used 15_J45014_F and Vr. We used a sample from each of our colonies, so that there were five GFP samples and five ATF1 samples. We also used a negative control for each of the two premixes and one positive control using the provided plasmid with Vf and Vr primers. We tried five different annealing temperatures (one for each sample) ranging from 48°C to 56°C - (the lowest melting point of our primers was 57.2°C).
- On Thursday we will run a restriction digest of all our samples, straight from the PCR.
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