Part:BBa_K676001:Experience

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Applications of BBa_K676001

According to our result in the 1D chloroquine gel electrophoresis (at 2.5 μg/mL), the plasmid with the Mu phage GBS (on pSB1C3 backbone) migrated further compared to the control plasmid (pSB1C3 with RFP expression cassette - BBa_J04450). At the chloroquine concentration which we used, the more negatively supercoiled plasmid will migrate further while the more relaxed molecules will travel slower through the gel.

The level of supercoiling achieved The plasmids with Mu GBS travelled further compared to the control . This indicated that the Mu GBS enables a higher supercoiled topoisomers to be formed . Overall ,the level of negative supercoiling ahcieved with the Mu SGS was

Calculations: By comparing the length of travel of the bands from the well in each samples

Control= 9.6cm

Mu GBS = 10.7cm

Difference: (1.1/9.6)(100) = 11.5%

Quality of supercoiling: Besides achieving a higher level of supercoiling, the quality of the supercoiled plasmids also showed to be better. The uniformity or homogeneity of the supercoiled Mu GBS plasmids increased by close to 30%( if the single distinct band was not included) compared to the control. Calculations: By comparing the range of topoisomers travel distances ( between the "slowest" and "fastest")

Control=4.9cm "smear"

Mu GBS =3.4cm "smear"

Difference: (1.5/4.9)(100%) = 30.6%

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