Difference between revisions of "Part:BBa J100175:Experience"
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Oligo Assembly: In one microfuge tube, we added 2uL (microliter) of 10X annealing buffer, 1uL of each of the 100 uM oligonucleotides (top and bottom), and 16 uL water to a total volume of 20 uL. The tube was boiled in approximately 500 mL of water for ten minutes and allowed to cool. Once cooled, we diluted the oligonucleotides one hundred fold and added 1 uL of the assembled oligos into the ligation. | Oligo Assembly: In one microfuge tube, we added 2uL (microliter) of 10X annealing buffer, 1uL of each of the 100 uM oligonucleotides (top and bottom), and 16 uL water to a total volume of 20 uL. The tube was boiled in approximately 500 mL of water for ten minutes and allowed to cool. Once cooled, we diluted the oligonucleotides one hundred fold and added 1 uL of the assembled oligos into the ligation. | ||
| − | [[File:partial j100175 | + | [[File:partial j100175 3.png]] |
This graph shows only the experimental promoter (J100175) and the negative control plasmid (J119137) to show the slight difference in values. | This graph shows only the experimental promoter (J100175) and the negative control plasmid (J119137) to show the slight difference in values. | ||
Revision as of 13:15, 2 October 2014
Oligo Assembly: In one microfuge tube, we added 2uL (microliter) of 10X annealing buffer, 1uL of each of the 100 uM oligonucleotides (top and bottom), and 16 uL water to a total volume of 20 uL. The tube was boiled in approximately 500 mL of water for ten minutes and allowed to cool. Once cooled, we diluted the oligonucleotides one hundred fold and added 1 uL of the assembled oligos into the ligation.
This graph shows only the experimental promoter (J100175) and the negative control plasmid (J119137) to show the slight difference in values.
This figure shows the experimental promoter (J100175), the negative control plasmid (J119137) and the positive control promoter (J04450). The positive control was known to produce Red Fluorescence, which accounts for the high values of fluorescence compared to the experimental and negative control.
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