1. Recombinant DNA Technology A set of methods used to locate, analyze, alter, study, and recombine DNA sequences Recombinant DNA is DNA in which nucleotide sequences from two different sources (even different species) are combined in the laboratory to produce a new combination of genes

2. Recombinant DNA Technology It is used to: –Probe the structure and function of genes If I delete this gene, what happens to the mouse? –Address questions in many areas of biology How are Bos taurus and Bos indicus related? –Create commercial products How can I get insulin production from bacteria? –Diagnose and treat diseases Does this child have cystic fibrosis?

3. Recombinant DNA Technology Using Mendelian and other genetic analyses we have talked about, everything we have deduced about genes and DNA sequence has been indirect With recombinant DNA technology we can isolate genes and DNA sequence, study them directly and store it in a convenient manner that facilitates future applications Cloning the DNA sequence accomplishes all of these

4. Review of steps involved in cloning http://highered.mcgraw- hill.com/sites/0072556781/student_view0/ chapter14/animation_quiz_1.htmlhttp://highered.mcgraw- hill.com/sites/0072556781/student_view0/ chapter14/animation_quiz_1.html

5. Recombinant DNA Technology Restriction enzymes were the key breakthrough in recombinant DNA technology and cloning Restriction enzymes are used to cleave DNA at specific sequences –Cuts DNA at specific sequences into different sized molecules –Cuts a single phosphodiester bond on each strand

7. Recombinant DNA Technology Recognition sequence –Usually four to six bases –Palindromes – read the same on both strands –Cut DNA reciprocally at specific recognition sites

9. Recombinant DNA Technology Most restriction enzymes make offset cuts –Restriction sites are not opposite each other Results in staggered cuts (sticky ends) Ability for hydrogen bonding to a complementary sequence