Agenda 11/9/16 Exam Unit 3 Intro to genetic engineering Activity: DNA Analysis Worksheet on Restriction Enzymes Learning Goal(s): Explain the role that restriction enzymes play in genetic engineering. Model the process of gel electrophoresis to analyze data commonly used in biotechnology. Describe the process of gel electrophoresis and it’s usefulness.

Genetic Engineering If you could genetically engineer something, what would it be and why would you do it? What would the benefit of your genetically engineered organism be? What are some ethical issues/cons regarding your engineered organism? Describe the process you would go through to engineer this organism.

Engineered Organism What would you engineer & why? What are the benefits of it? What are the possible issues/ethics involved? Describe the process

Genetic Engineering The Manipulation of DNA What can it be used for?

Manipulating DNA for… Studying Genealogy Solving crimes Studying/Curing disease (gene therapy) Pure science (knowledge) Studying Evolution (changes in allele frequency, resistant strains of bacteria) Advances in agriculture (GMOs) ????

How do we manipulate the DNA? What techniques do we employ?

Review DNA Analysis by Gel Electrophoresis Gel electrophoresis is a technique used for separating DNA fragments for analysis. DNA moves through a gel (agarose) when an electric field is applied because DNA is negatively charged. DNA fragments are separated by size in the gel. Used in forensics, genetics, microbiology, biochemistry, & molecular biology.

How is DNA Manipulated for Gel Electrophoresis? DNA has to be cut to produce fragments. Restriction Enzymes cleave (cut) nucleotides at specific sites. Some leave “sticky ends”, others “straight ends”. Examples of restriction enzymes: HaeII (straight cut) Bam EcoRI, EcoRII HindII (straight cut), HindIII Different lengths of DNA are produced from the cutting process 3. Those fragments are then separated using gel electrophoresis technique Shorter fragments will “run” farther away from origin Longer fragments will separate out earlier 4. Analysis of DNA is based on the banding pattern

Example Original Strand of DNA: ACCGGTGA TGGCCACT After digestion (cut) with restriction enzyme: ACC GGTGA TGG CCACT Can you tell what the cut site was? Remember the palindromes? The results are different lengths of DNA sequences (called fragments). Next the fragments are arranged from largest smallest fragment size (these are the different bands you see on the gel). GGTGA ACC CCACT TGG

DNA Fingerprinting Analysis Gel ran from top to bottom. Where would the longer fragments be located? There are six lanes of DNA fragments Which two lanes have matching DNA?

Who should pay child support? How do you know? The lover because bands 4 and 8 match up only with him. Source:http://www2.ups.edu/faculty/kirkpatrick/bio111/studyquestions/00answers/12adna%26rep%26technology1.htm

Who is the perpetrator? Suspect 2 because the DNA patterns are the same. Source:http://www2.ups.edu/faculty/kirkpatrick/bio111/studyquestions/00answers/12adna%26rep%26technology1.htm

Restriction Enzyme Activity Cut the 5 segments of DNA (1 from evidence collected at a crime scene plus the 4 suspects) ONE AT A TIME. The restriction enzyme (HaeIII) recognizes the CCGG sequence and cuts the double helix between the C and G. Example: ACC GGTGA TGG CCACT The results are different lengths of DNA sequences (called fragments). Next, glue the fragments from each sample onto blank paper from largest smallest fragment size. Example: GGTGA ACC CCACT TGG Be sure to label the DNA fragments with the appropriate name (ie. Crime, Suspect 1, Suspect 2, etc…) Put a star * next to the suspect’s DNA that matched the crime scene DNA.

Review What is used to cut DNA into fragments? Restriction Enzymes What technique is used to separate those fragments? Gel electrophoresis Which fragments separate out the fastest? Longest fragments Which fragments separate out the slowest (because they “run” farther)? Shortest fragments What can the banding pattern tell us? Size of fragments relatedness

Restriction Enzyme W.S. Complete the worksheet and turn it in.

Pick one of the Crime Cards Pick up the crime background sheet and read it. Pick up 2 DNA evidence cards, and a suspects DNA card. Compare the cards to determine who the perp is.

Video: genetic engineering microbes https://www.youtube.com/watch?v=zlqD4UWCuws Genetic Engineering for Human Enhancement. https://www.youtube.com/watch?v=mXfYshYnbIA Nova: Crispr article http://www.pbs.org/wgbh/nova/next/evolution/crispr-gene-drives/

With a Partner…. Make a list of possible applications for genetic engineering and potential problems that may be solved using this technology. After watching the video clips, add to your list. Add a list of potential problems/issues/remaining questions that may arise as a result of this technology. Discovery streaming : How stuff works (wheat, segments 7 & 8) How stuff works: Super Trees (timber, segment 3)