The primer sequences on the PCR products each have an EcoRI restriction enzyme site. Your plasmid contains an ampR gene and a b-galactosidase (lacZ) gene. The lacZ gene contains an EcoRI restriction enzyme site. After you digest the PCR products and your plasmids with the EcoRI restriction enzyme explain the following:

The lacZ gene from coli encodes the enzyme beta-galactosidase, which can catalyze the conversion of a colorless compound called X-gal into a blue product. Molecular biologists have taken advantage of this property by constructing plasmid vectors that contain the lacZ gene with an EcoRI restriction site in the middle of the coding region.

The restriction enzyme EcoRI recognizes the DNA sequence 5’-GAATTC-3’. The “cut” in the phosphodiester backbone happens after the first G. Draw (using letters, not chemical structures) both strands of the DNA after cleavage of the recognition sequence (include the terminal phosphate group and terminal –OH.)

The results are as follows: Restriction Enzyme(s) Fragment lengths (base pairs) EcoRI BamHI HindIII EcoRI and BamHI30 bp, 70 bp, 305 bp, 400 bp EcoRI and Hindill BamHI and HindIlI105 bp, 230 bp, 470 bp 430 bp, 375 bp 470 bp, 335 bp 805 bp 75 bp, 300 bp, 430 bp Practice Final

Assume you cut this fragment with the restriction enzyme EcoRI. The restriction site for EcoRI is 5’-GAATTC-3’, and the enzyme makes a staggered (“sticky end”) cut between G and A on both strands of the DNA molecule. Based on this information, draw the cuts on the DNA sequence above by highlighting the target sequences.

Question: _____ is a restriction enzyme. _____ is DNA. pBR322, calf thymus DNA EcoRI, BamHI EcoRI, pBR322 RNA, EcoRI

The EcoRI restriction enzyme has the recognition sequence/site seen below. If it were used to cut the (linear) sequence of DNA also shown below: (Total 2 points) a. How many fragments would be produced?

Question: 1. The restriction enzyme EcoR1 cuts DNA at the following sequence: 5'-GAATTC-3' 3'-CTTAAG-5" Assume you use EcoRI to digest this strand of DNA: 5 ...

Question: You have a circular plasmid that is 5,000 bp long. After treating it with the EcoRI restriction enzyme, you loaded the plasmid DNA in an agarose gel and ran gel electrophoresis. The results of the gel show two bands at the 2,000 and 3,000 bp marks on the ladder. How many EcoRI cut sites were present in the plasmid?

After cutting the plasmid with the above three restriction enzymes, you decide to try another restriction enzyme, EcoRI. Based on the results of your gel, what is your interpretation of the DNA bands you observe in the EcoRI lane? ( 0.5 points)I see that the EcoRI has two recognition sites and is about 5200 base pairs in length.