1. Applications and Basic Technology

2.  Recombinant DNA technology : set of techniques for recombining genes from different sources and transferring into cells where it may be expressed

3.  Occur in bacteria where they protect against intruding DNA  Involves restriction; foreign DNA is cut into small segments  Cut at specific points on small segments called specific recognition sequences  Several hundred restriction enzymes & ~ 100 different specific recognition sequences  Cuts DNA in staggered manner, resulting in single-stranded ends, called sticky ends

7.  Creates restriction fragments  Used in lab to join DNA pieces from different sources  Temporary bonds held by weak H bonds  Can be made permanent by adding DNA ligase

8.  used to separate either nucleic acids or proteins based upon molecular size, charge and other physical properties  used to identify segments by looking at banding pattern when cut with restriction enzymes  DNA fragments can be isolated, purefied, and then recovered from gel

9.  Electrical current is used to move charged particles at different rates

11.  gel electrophoresis of restriction fragments results in characteristic banding pattern  Each band corresponds to DNA restriction fragments of certain length  Different alleles of gene result in dissimilar banding patterns  Similar patterns result when noncoding segments of DNA are used as starting materials

12.  Fragments move different distances based on base pair length, size (how compact)

13.  Used to match DNA from different sources

14.  Use restriction enzymes to alter DNA  Incorporate DNA into new organism to create a GMO (genetically modified organism)  Ex: Super Salmon, Flavr Savr Tomato  Super Salmon ABC Special Super Salmon ABC Special

15.  Technique used to amplify (copy many times) DNA in vitro (inside the cell)  1. DNA is incubated with special primers & DNA polymerase  2. billions of copies of the DNA are produced in a few hours  3. PCR is highly specific; primers determine sequence to be amplified  4. only minute amounts of DNA are needed