S-STEM Scholar Leilani Boren's Blog

 Introduction: The previous week resulted in a gel that revealed 54 degrees annealing temperature for overlap PCR reaction 1, the reaction that is without primers responsible for binding the two homologous regions of left and tetr/a fragments, is the most efficient method for two way assembly product so far. Since 54 degrees Celsius was the highest temperature tested in the gradient experiment, it was decided to evaluate even higher temperatures to be able to fully visualize the full range of possibilities for optimal two way assembly with minimal ghost band products and high yield of desired product. The remaining amount of sample was loaded onto a gel for clearer visualization of last week's results, and then a higher heat gradient was conducted with left and tetr/a for the final time. Once a temperature is decided, the project can move forward by isolating the left and tetr/a overlap product so final three way assembly with the addition of the right fragment can be attempted in ...

 Introduction: The previous weeks have led to confusion surrounding the ideal temperatures for annealing PCR conditions The most recent gel which included a 34 to 54 degrees Celsius heat gradient to evaluate the optimal temperature for left and tet fragment assembly resulted in a gel with no DNA visible and no primers. There was suspicion surrounding a mixup of tubes during that process which would explain the lack of DNA, since that result had not happened before amongst all of the trials this semester. A preliminary test was done amplifying another set of tubes which could have been mixed up with the tubes actually amplified the previous week, to check for any results. Afterwards, this week we carried out a re-do of the previous week's experiment and got partial clarity for steps moving forward.  Methods: Due to the suspicion of two sets of eight tubes being confused with the other set of eight, which were amplified and ran to reveal no gel results, the tubes labeled "Overla...