Molecular Biology Summer 2021 Notebook

6/15/21

Maxiprepped SARS-CoV-2-S-RBD-sfGFP_pcDNA3-1, SARS-CoV-2-S-RBD-sfGFP_pcDNA3-2, hACE2_pcDNA3.1-1, and hACE2_pcDNA3.1-2. Left to dry after ethanol step.

6/14/21

Ran and analyzed gel for SARS-CoV-2-S-RBD-sfGFP_pcDNA3-1 and 2 and hACE2_pcDNA3.1-1 and 2.

6/10/21

Resuspended KCNJ3-1 and KCNJ3-2. Ran KCNJ3-1, KCNJ3-2, OPRM1-1, OPRM1-2, mRuby3-1, mRuby3-2, KCNJ9-1, and KCNJ9-2 on a gel at 160V for 45 minutes. Inoculated SARS-CoV-2-S-RBD-sfGFP_pcDNA3 twice and hACE_pcDNA3.1 twice.

6/09/21

KCNJ3-1 and KCNJ3-2 were spun in the centrifuge at 13,000g for 5 minutes in order to obtain a pellet. A pellet was obtained, and 70% ethanol was added. They were then spun at 10,000g for 2 minutes. They were left overnight with the caps off so that the 70% ethanol would evaporate. Resuspended OPRM1-1, OPRM1-2, KCNJ9-1, KCNJ9-2, pKK-mRuby3-TEV-1, and pKK-mRuby3-TEV2. SARS-CoV-2-S-RBD-sfGFP_pcDNA3 and hACE2_pcDNA3.1 were streaked for single colonies.

6/08/21

Maxi-prep of OPRM1, KCNJ3, KCNJ9, and pKK-mRuby3-TEV. Stopped at step 8 for KCNJ3-1 and KCNJ3-2 as no pellet had formed after centrifuging at 10,000g for 5 minutes. 1/10 of the estimated volume in the tubes of 3M sodium acetate buffer was added and the tubes were left to sit overnight with the caps on. OPRM1, KCNJ9, and pKK-mRuby3-TEV were left overnight with the caps off so that the 70% ethanol would evaporate.

6/07/21

Gel Electrophoresis of OPRM1, KCNJ3, KCNJ9, pKK-mRuby3-TEV was done to determine the length and concentration of the DNA and the accuracy of the maxiprep. Transformation of pKK-mRuby3-TEV. No page was made for this transformation.

6/03-04/21

Maxiprep of OPRM1, KCNJ3, KCNJ9, and pKK-mRuby3-TEV was performed. The Maxiprep failed and took multiple days with the freezing of bacteria and DNA in the middle due to shortages of solutions.