Part:BBa_K2607002:Experience

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Experiment with BBa_K2607002 and BBa_K592024

We performed experiment to compare BBa_K2607002 with BBa_K592024. Figure 1 shows the plan on how we conduct the experiment.

Figure 1. Original plan workflow with newly-established (BBa_K2607002) and existing (BBa_K592024) parts of BFP.

Gel Electrophoresis Confirmation
Upon receiving our “Improved BFP” gBlock from IDT (with length of 1151 basepairs (bp) with SalI and NdeI restriction sites are located at base 244-249 and 266-271, respectively), we performed PCR to amplify it until sufficient quantity (about 100 ng/uL). We then created two restriction digestion reactions, one with NdeI restriction enzyme and one with SalI restriction enzyme. Each reaction comprised of 100 uL PCR-amplified “Improved BFP” gBlock, 6 uL restriction enzyme, 15 uL CutSmart® restriction buffer, and 29 uL nuclease-free water. The reaction was subsequently incubated for four hours at 37oC incubator. The digestion products were then run for electrophoresis in 1% agarose gel with tris-acetic acid-EDTA (TAE) buffer 1x, power supply 50 volt for 70 minutes. Visualization was performed with Bio-Rad Gel DocTM XR+ Gel Documentation System.

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