Lab 7 Gene Cloning Plasmids and a Restriction Endonuclease
Chang Mo Yeon # 100777264 March-13-13 Friday P.M
Figure 1. The standard curve of log of the base pair of the bands from the ladder used for agarose gel versus the mobility of the protein bands eluted
Figure 2: Image of the DNA agarose gel electrophoresis: undigested and digested lamada DNA, standard pBR 322, standard pUC18 and undigested and digested of sample 1 and 2. Original bands are labeled (1 – 29) and expected bands are also labeled. Table1: Identification of DNA agarose gel electrophoresis bands.
Band # 1 2 3 4 5 6 7 8 9 10 Identification intact λDNA genome intact λDNA genome λDNA fragments λDNA fragments λDNA fragments λDNA fragments λDNA fragments λDNA fragments pBR322 cccDNA dimer pBR322 cccDNA Band # 11 12 13 14 15 16 17 18 19 20 Identification pBR322 ocDNA pUC18 ocDNA pUC18 cccDNA pUC18 ocDNA λDNA fragments pBR322 ocDNA pUC18 ocDNA pUC18 cccDNA pUC18 cccDNA λDNA fragments Band # 21 22 23 24 25 26 27 28 29 Identification pBR322 ocDNA pUC18 ocDNA pUC18 cccDNA λDNA fragments pBR322 ocDNA pUC18 cccDNA pUC18 cccDNA pBR322 ocDNA pUC18 cccDNA
Column explaining the conformation of plasmid: λ DNA undigested should only show 1 band because plasmid is all pen. Undigested puc 18 shows 3 bands: 12, 13 and 32. Respectively nicked and supercoil and liner are conformed. Supercoil tends to be the brightest band and nicked are the slowest because they are circular plasmid and very relaxed and they normally replicates. Same goes to pBR 322 nicked, supercoil and linear for bands 9, 10 and 31 respectively. Supercoil are the fastest that migrates down the gel. Sample 2 digested only produced supercoil with the bright band (28), where as sample 1 undigested and digested and sample 2 undigested produced more band.
Table 2. Identification of DNA agarose gel electrophoresis bands of expected and missing bands.
Band # 30 31 32 Identification λDNA fragments pBR322 cccDNA pUC18 cccDNA
Table 3. DNA...