October 5, 2019
We started our lab session by first collecting our tubes of Mimulus guttatus DNA. We obtained 12 1μL tubes and labeled them ‘QS 1-12.’ QS stands for our group name ‘Queen Salmon.’ We were sure to write down the corresponding specimen ID number to the tube number assigned to each group member.
We then proceeded to make the master mix that was a key part of this PCR reaction. We were given the amount of each ingredient that was made for one reaction. Since we had 12 PCR reactions to complete, we decided to multiply these values by 18 to account for each reaction as well as any error that could occur. The ingredients consisted of; 240.48μL ddH2O, 36μL 10x buffer, 36μL MgCl2, 18μL BSA, 3.6μL dNTPs, 3.6μL F-primer, 3.6μL R-primer, 0.8μL Taq. This mixture was then vortexed to make sure that the ingredients were properly mixed together.
We then took the new labeled tubes and added 19μL of the master mix as well as the 1μL template from our individual samples. The tubes were then vortexed for a few seconds before being stored on ice.