User:Daniel Ramirez/Notebook/UNAM Genomics Mexico 2011/2011/07/12

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χρόνος πέρασμα July 12th 2011


  • Pamela Silver Lab's plasmid digestion with XbaI. This digestion was performed just to see if the DH5α E.coli indeed carry these plasmids.

  • Today I made gel electrophoresis to see if Pamela Silver Lab's plasmids were correctly digested with XbaI. What I expected to see was the following:
  • The plasmid pPS3182 that contains HydEF and HydG is 3781 bp long without any gene cloned into its multiple cloning sites; as this plasmid contains two genes cloned (HydEF and HydG into MCS1 and MCS2, respectively), each standarized as a biobrick, and as there's one XbaI restriction site inside the backbone of the plasmid, three fragments are expected if the digestion was performed correctly. Fragment #1 - 1500 pb, fragment #2 - 3756 bp, fragment #3 - 3770 pb.
  • The plasmid pPS3183 that contains PFOR (cloned into MSC2) is 4008 pb long without any gene cloned into any of its two multiple cloning sites. This plasmid contains only one biobrick standarized gene, and there is one XbaI restriction site in the plasmid backbone; therefore, two fragments are expected if the plasmid has been properly digested. Fragment #1 - 1715 pb, fragment #2 - 6091 bp.
  • The plasmid pPS3186 that contains HydA and Fd is 5420 bp long without any gene cloned in it. Tha plasmid contains two biobrick standarized genes (HydA and Fd into MCS1 and MCS2, respectively), no other XbaI site is found in the plasmid backbone. Two fragments are expected as the digestion outcome. Fragment #1 - 1951 bp, fragment #2 - 5431 bp.
  • Here's the gel that shows all three plasmids correctly digested. Well #1 and #6 contains 1kb ladder, well #2 contains plasmid pPS3182, well #3 contains plasmid pPS3183, well #4 contains plasmid pPS3186 and well #5 contains only dye as a negative control.

PSplasmids XbaI digestion.jpg