On Monday and Tuesday I performed PCR reactions and cloning prep to eventually sequence mouse umi DNA. I had to run PCR twice to amplify DNA and used different primers each time. In the first reaction I had a positive and negative control, with a different primer and water respectively, in order to make sure that my results are successful not because of human error. The second reaction is called a nested reaction which is used as the final reaction before gel purification.
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I then performed gel purification. This is how labs isolate DNA. I had to go into a room with a Plexiglas hood mask and flame resistant lab coat to protect myself from the UV light. Using a sharp knife and a UV wand for illumination, I cut out the two visible DNA bands and allocated them to two tubes. Next, I prepared for plasmid bacterial cloning and later MiniPrep. This process is used to add more sequenced data to the Mouse and Human database for analysis on T Cell biases.
On Wednesday, I went to a Giants baseball game with a couple members of my lab. This was probably the best game to go to because it was a no hitter game (a rare occurrence in baseball). Thus, the intensity was high throughout the game and it was a great lab bonding experience.
On Thursday I finished my mini prep sequencing and on Friday my PI gave me rat primers which I will be using to perform TCR (tell cell experiments) for rat T cells. I had to dilute and allocate these primers, carefully calculating the amount of water needed. On Monday I am going to dissect dead rats to remove the organs containing high amounts of T Cells. Later I will extract blood (serving as template DNA) for my PCR reactions.
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