Week One: 1/22-1/26 Growth and Extraction of the pRHAM Plasmid
Monday On Monday, I started out the week by preparing LB Broth and LB Broth with Agar. I made LB media because the pRHAM plasmid is extracted from E.Coli which can only be grown on LB. So, I made 200 mL of LB Agar to prepare around 8 plates once autoclaved and heated, but that will be touched on in a later day. Along with the LB Agar I had also prepared 250 mL of LB Broth which will be used to make smaller flasks of LB to grow the E.Coli in. Once both of those broths were prepared I put them on the autoclave tray and waited for them to be autoclaved for use the next day. Before I left for the day however, I calculated how much kanamycin I needed for both the broth and the broth with agar. I did this by getting the stock concentration that we had, which was 7.5 mg/mL, finding the desired concentration which was 30 ug/mL and then using the desired volume which was either 200 mL for the agar or 250 mL for the broth without agar. The reason kanamycin was used is because that specific ...