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

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

4. Palindromes:Example of Restriction Sites:

5. 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 12.09.10:

6. 2300400030005000bp 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.

7. 23000 bp 9000 bp 6000 bp 4000 bp 2000 bp 500 bp 2300bp 2300400030005000bp E E+H EHB H B EcoRI HinDIII BamH1

8. 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.

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

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

11. 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 4720-702=4018bp Expected fragments

12. 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

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