Experiments

The design of DNA fragment (Seq.1) we used this year is shown below. In PURE system, T7 RNA polymerase is contained. Basically, we need to use T7 promoter designated by the inventors for transcription. SD sequence and Red fluorescent protein lie between BBa_K1859015 and BBa_K1859016. As terminator of this alignment, T7 terminator (Bba_K731721) was selected. In case of myTXTL, it uses σ70 promoter. When you use T7 promoter, you need to add T7 RNA polymerase. The recommended concentration of the polymerase is 2U/μL. SD sequence (RBS) and RBS is sandwiched by ribosomes. As T7 promoter, we did not use the sequence which has been registered in iGEM catalog. Instead of this, we used the new sequence of T7 promoter.