Before beginning the protocol for the October 24, 2018 lab, we ran gels of the three DNA plant samples we extracted the week before. The gels were loaded with the samples of an entire lab table. The loading dye was pipetted onto a strip of filiminet paper in clusters of three dots for each sample the four members need to load. An image of the order is included below. The samples were pipetted onto the dye and transferred into the gel wells. Once the gel was run for 20 minutes it was imaged and the picture was saved to use in future blog write-ups. Our gel presented 12 samples that migrated across the gel. Following the gel procedure, 20μl of each of my three samples were transferred into two new tubes. The two new tubes for each sample was labeled with its corresponding sample ID. The samples were distributed between the three tables of four students in the class, so each student was responsible for 8 different samples of Mimulus cardinals DNA.

The plant DNA PCR- EPIC markers protocol guided us through the preparing of the template DNA. One 0.2ml 8-strip of PCR tubes was labeled with the 8 different samples IDs assigned. Each tube was labeled with the primer type as well. In our case we used EPIC 5525. Into each tube 1μl of template DNA was dispensed using a filter tips and a pipet. Each sample was placed into its corresponding tube, and different filter tips were used for each template DNA. Then, the strips were sealed and placed on ice.

Setting up the Master Mix was next on the protocol. The amount of reagents needed for the table was calculated for 40 samples. A 1.5ml tube was labeled MM for Master mix and set on the rack. The reagents pipetted into the tube included: 60μl of ddH2O, 80μl of 10x buffer +Mg, 40μl of BSA, 8 μl of dNTPs, 8μl of F-primer, 8μl of R-primer, and 1.6μl  of Taq. While one member created the master mix, the others transposed the IDs on their tubes onto the PCR sheet as seen in the image below. 19μl of Master Mix was pipetted into all 8 tubes using different tips for each tube. The content was mixed using the pipet. Finally, the strip of tubes was sealed and place in the PCR machine and the lid was closed.

Image Of Loading Gel Order: 

Image of the Calculated Reagents and 8 Specimen ID: