Recognition sequences Restriction enzyme EcoRI cuts the DNA into fragments. Sticky end

Originally protected bacteria from viruses. Restriction Site = Recognition sequence NOT Cleavage site Recognition site is a planidrome (just like “radar” “Madam, I’m adam”

Palindromes:Example of Restriction Sites:

1. Which sizes of DNA fragments do you expect to get from the digestion? 2. How can one use the electrophoresis procedure to estimate the size of the DNA fragments? Journal :

bp EcoRIHinDIIIBamH1 1.Which fragment sizes will result from cutting with: A. HinDIII alone? B. EcoE1 alone? C. Both HinDIII+EcoR1 2. Draw the sizes into the gel.

23000 bp 9000 bp 6000 bp 4000 bp 2000 bp 500 bp 2300bp bp E E+H EHB H B EcoRI HinDIII BamH1

To Students that are presenting about restriction maps and expected sizes: You may want to peak through the slides about gel electrophoresis, to see the connection between your part and the next – electrophoresis.

Restriction digest of pARA-R Recombinant plasmid of interest pARA-R BamH I Hind III rfp 702bp 4720 bp

pARA-R construct Recombinant plasmid of interest pARA-R BamH I Hind III rfp 702bp 4720 bp rfp 702bp pARA-R 4720 bp

pARA-R construct Recombinant plasmid of interest pARA-R BamH I Hind III rfp 702bp 4720 bp rfp 702bp pARA-R 4720 bp 702 bp =4018bp Expected fragments

restriction analysis of pARA-R BamH I Hind III 702bp BamH I Hind III Restriction fragments after digest with Hind III and BamH I 4018 bp

Additional Resources: Restriction Map Exercise for high-school (if you want to use this)