Nt1310 Lab 9

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The following results helped obtain the haplogroup that in which the sequence of mtDNA would identify. The PCR reaction worked, and this can be determined by looking at the agarose gel in figure 1. If the PCR reaction was successful, than a band should appear around 550bp. Individual AC displays a band around 550bp, this means the PCR reaction was successful. The band for individual AC, depicts a low concentration of product, because the band faint. After the purification process the concentration, A260/280 ratio, and A260/A230 ratio were determined by using the nanodrop. The concentration of mtDNA in the product was 60.9 ng/uL. The ratio for A260/280 was 1.79 and the ratio for A260/230 was 0.77. The A260 and 280 are a spectrometer measurement that measure absorbance at wavelengths of…show more content…
Therefore the ratio of A260/230 is the concentration of DNA/ the concentration of other containments (Wilfinger). From these values one can determine that the sequencing process was successful. These values displayed that the sequencing reaction worked, but did not give the desired results and did not have a concentrated product. The amount of the DNA sample and water had to be adjusted in the tube that was going to be sent to the Cornell lab. The amount that was supposed to be used was 1.5ul of mtDNA. This value was determined by 100ng of DNA required x volume of DNA in ul divided by the concentration of DNA. The instructor simply advised to double the amount of mtDNA that would be placed in the tube. The volume of DNA used was 3uL, and the volume of sterile water used was 13.4uL. The volume of the primer used was 1.6uL(which was not adjusted). Figure 2 displays the reverse complement of the mtDNA that was obtained from the Cornell lab sequencing the mtDNA sent to them. This was then used to compare it with the CRS mtDNA. Figure 3 displays the aliment of the CRS and the revers compliemt of the mtDNA sent to the Cornell
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