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A Flower of A Different Color
We, vector group (Angie and Diwash) did the isolation of plasmids pSB1A3 and pSB1A7 by using the 'Gene jet plasmid miniprep kit" from the Fermantas company. We also prepare the agarose gel to run the plasmid samples. After the plasmid isolation, we did the single restriction digest of each plasmid samples by using the fast digest SpeI and Xbal. After the plasmid isolation, we run the plasmid samples in the agarose gel for 20 minutes at 150 Volt. The picture from the gel shows the plasmid bands of around 2100 bp and around 1700 bp when digested with Xbal and SpeI in pSB1A3 and pSB1A7 respectively. As, pSB1A3 should be 2157 bp and pSB1A7 should be 2431bp. Hence, from the experiment it is clear that pSB1A7 is not the correct plasmid for us to insert the cytochrome p450 gene. Instead, the result from pAB1A3 proves that it is a correct plasmid to insert our gene of interest. Also it varifies that somebody has switched the vector during the experiment 15,hence the result was just opposite at that time.
Sushma and Binu did the transformation of DH5alpha cells with each of the ligated samples of 11/17/2008 and incubated them overnight at 37 degrees.