Amplification of Potential Past 2-Way Assemblies
Traditional PCR of Potentially Assembled Fragments October 28th, 2022 Introduction In hopes of running into a 2-way assembly product from all past Overlap PCRs, this procedure aims at amplifying the Left+Kanamycin and Right+Kanamycin products using traditional PCR. Over the past few months, multiple Overlap Extension PCRs were carried out either by the mentors Shawn and AJ or by myself after I took over the project. Since one Overlap Extension PCR was successful in the past, this procedure aims to point to the assembly products that, hopefully, were not lost or used up in other procedures. It is hypothesized that at least one Left+Kanamycin product and one Right+Kanamycin product will show a favorable band on the gel. Procedure Using old tubes labeled LM (left+middle) and RM(right+middle) as the template DNA because they were the assembly products of Overlap PCR, the amplification of the Left+Kanamycin fragments of the assembly products will utilize primers 1 (forward)...