![]() For some samples, three bands are obtained from the DNA after digestion, the three bands are at about 1500-3000 bp location, it showed that the DNA are not completely digested. Therefore, the first band should be ignored when considering BBa_J04450.įor all our DNA samples, they have at least two bands for DNA, whereas the two bands showing that the DNA is digested. The open circular BBa_J04450 has a smaller electrophoretic mobility, so it moves slower, and migrates less distance.įor our experimental DNA samples, most of them showed four bands, in which the first band all have the same number of base pair, which are higher than 3000 base pair, showing that this may be an unexpected DNA fragment, as the number of base pair of BBa_J04450 is 3139 bp. Since DNA ladder is smearing, it is not helpful to identify the size of DNA samples.įrom the negative control, the linear BBa_J04450, it shows two bands, which may be caused by the presence of other DNAs, but not solely BBa_J04450. To find out whether 200ng of DNA can be digested by using 0.2 μL of restriction enzyme in 5, 15 and 30 minutes. NotI-HF restriction enzyme site in pSB1C3 from NEBcutter V2.0 BBa_J04450 plasmid with XbaI restriction enzyme site from iGEM Registry. We choose to use linear DNA for this investigation, which is BBa_J04450.įigure 2. However, it is found that in NEB website, it claims that there are some enzymes which are qualified for TimeSaver™ can digest DNA for only five minutes, thus we would want to investigate on the shortest time that restriction enzyme can digest DNA so to reduce time cost for future researches. For normal practice, researchers digest DNA using restriction enzyme for 1 to 1.5 hours.
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