Model of DNA DNA is Comprised of Four Base Pairs

1. Model of DNADNA is Comprised of Four Base Pairs

2. DNA Restriction Enzymes • Evolved by bacteria to protect against viral DNA infection • Endonucleases = cleave within DNA strands • Over 3,000 known enzymes

3. Enzyme Site Recognition Restriction site Palindrome • Each enzyme digests (cuts) DNA at a specific sequence = restriction site • Enzymes recognize 4- or 6- base pair, palindromic sequences (eg GAATTC) Fragment 2 Fragment 1

4. 5 vs 3 Prime Overhang Enzyme cuts • Generates 5 prime overhang (Extremos 5’ salientes) 5’GGGCCC3’ 3’CCCGGG5’ 5’GGG CCC3’ 3’CCC GGG5’ Extremos romos

5. Common Restriction Enzymes EcoRI – Eschericha coli – 5 prime overhang Pstl – Providencia stuartii – 3 prime overhang 5’CTGCAG3’ 3’GACGTC5’ 5’CTGCA3’ G3’ 3’G ACGTC5’ Extremos 3’ salientes

6. DNA Fingerprinting Real WorldApplications • Crime scene • Human relatedness • Paternity • Animal relatedness • Anthropology studies • Disease-causing organisms • Food identification • Human remains • Monitoring transplants

7. Workshop Time Line • Restriction digest of DNA samples • Introduction to DNA Fingerprinting and RFLP analysis • Electrophoresis on Agarose gels • Analysis and interpretation of results

8. DNA FingerprintingProcedureOverview

9. DNA Fingerprinting ProceduresDay One

10. DNA Fingerprinting ProceduresDay Two

11. DNA Fingerprinting ProceduresDay Three