Final Review The following slides are slides we used to prepare for the DNA Technologies quiz. Review the sites and materials listed to help prepare you for the final. Your worksheets from this section will be valuable as study aides as well. There are also some questions at the end we used previously to prepare for this section’s quiz.

Types of Cloning 1 recombinant DNA technology or DNA cloning, the transfer of a DNA fragment of interest from one organism to a self-replicating genetic element such as a bacterial plasmid. The DNA of interest can then be propagated in a foreign host cell. 2 reproductive cloning Reproductive cloning is a technology used to generate an animal that has the same nuclear DNA as another currently or previously existing animal. In a process called "somatic cell nuclear transfer" (SCNT), genetic material from the nucleus of a donor adult cell is transferred to an egg whose nucleus, and thus its genetic material, has been removed. 3 therapeutic cloning "embryo cloning," is the production of human embryos for use in research. The goal of this process is not to create cloned human beings, but rather to harvest stem cells that can be used to study human development and to treat disease.

Applications 1 recombinant DNA technology or DNA cloning, This type of cloning is important for learning about other related technologies, such as gene therapy, genetic engineering of organisms, and sequencing genomes Gene therapy can be used to treat certain genetic conditions by introducing virus vectors that carry corrected copies of faulty genes into the cells of a host organism. Genes from different organisms that improve taste and nutritional value or provide resistance to particular types of disease can be used to genetically engineer food crops. 2 reproductive cloning Reproductive cloning also could be used to repopulate endangered animals or animals that are difficult to breed 3 therapeutic cloning Therapeutic cloning technology may some day be used in humans to produce whole organs from single cells or to produce healthy cells that can replace damaged cells in degenerative diseases such as Alzheimer's or Parkinson's.

Stem cells—Cells with the ability to divide for indefinite periods in culture and to give rise to specialized cells. Embryonic stem cells—Primitive (undifferentiated ) cells derived from a 5-day preimplantation embryo that are capable of dividing without differentiating for a prolonged period in culture, and are known to develop into cells and tissues of the three primary germ layers. Somatic (adult) stem cells—A relatively rare undifferentiated cell found in many organs and differentiated tissues with a limited capacity for both self renewal (in the laboratory) and differentiation. Such cells vary in their differentiation capacity, but it is usually limited to cell types in the organ of origin. This is an active area of investigation. Induced pluripotent stem cells—Somatic (adult) cells reprogrammed to enter an embryonic stem cell–like state by being forced to express factors important for maintaining the "stemness" of embryonic stem cells (ESCs). Scientists are actively comparing iPSCs and ESCs to identify important similarities and differences.

Adult Vs. Embryonic One major difference between adult and embryonic stem cells is their different abilities in the number and type of differentiated cell types they can become. Embryonic stem cells can become all cell types of the body because they are pluripotent. Adult stem cells are thought to be limited to differentiating into different cell types of their tissue of origin.

Cloning Continued Today you will complete the “Click and Clone” activity from the Utah Genetics web page Answer the questions provided on the worksheet as you try your hand at cloning for yourself. Once you are done turn in the worksheet and review the information provided in your packet on “DNA sequencing”. Answer the following questions about this process What can the sequence of a gene be used to predict? In sequencing what will cause the synthesis of a new strand to stop? What is the result of this halt in strand synthesis? How do we determine the sequence of the original DNA template at the end of the reaction? Have the answers to these questions completed on a sheet of notebook paper for Monday

Sequencing Now that you have read a bit on the process of sequencing we will take a look at the steps of this procedure http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/D/DNAsequencing.html Review the information on this web page concerning the steps of sequncing . There are many steps so you may want to take some notes to help you keep track of this process. Q: What is the difference between normal (deoxy) nucleotides and dideoxynucleotides Q: What is the other name for the dideoxy method? Why? Once you have reviewed this web page and answered the questions above, click on “Animation of this proccedure”. Answer the questions provided on the worksheet as you go through the steps of DNA sequencing.

DNA Technologies The Genetic Science Learning Center Cloning, Sequencing and DNA Microarrays 1. Cloning Article Analysis TEXT= 366 to 368 2. Cloning techniques, recombinant DNA cloning animation TEXT= 373, 367and 368 3.DNA Microarrays TEXT= 375to 379 Review your cloning knowledge The Genetic Science Learning Center http://learn.genetics.utah.edu/content/tech/cloning/whatiscloning/ Read over the steps of somatic cell nuclear transfer (SCNT) and then view the two animations comparing Natural reproduction to SCNT. Summarize the difference between Natural reproduction and SCNT Describe what is done to the cell called the “ egg cell donor” in SCNT

Review http://www.dnalc.org/resources/animations/cycseq.html Topics we have covered include: Cloning types and techniques Sequencing steps and results Microarray procedure and application DNA Dragnet use and social impact Plasmid Cloning Methods Investigation http://www.sumanasinc.com/webcontent/animations/content/plasmidcloning.html “Cycle Sequencing” on Dolan DNA Learning Center http://www.dnalc.org/resources/animations/cycseq.html http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/D/DNAsequencing.html Cloning Article responses http://www.ornl.gov/sci/techresources/Human_Genome/elsi/cloning.shtml http://stemcells.nih.gov/info/basics/basics1.asp

“Click and Clone” on Utah Genetics http://learn.genetics.utah.edu/content/tech/cloning/ “Sequence For Yourself” http://www.pbs.org/wgbh/nova/genome/sequencer.html#

DNA technology review questions Answer the following questions to the best of your ability. On Monday we will take a look at each of the techniques we have covered and the steps involved in these processes.

1. Which of the technologies listed below is a valuable method for mass-producing drugs and other useful proteins? A. B. C. D. recombinant DNA technology transgenic technology biotechnology gene targeting 2. Manufacturing recombinant DNA molecules involves cutting a gene from its normal location, inserting it into a circular piece of DNA from a bacterial cell, and then transferring the circle of DNA to cells of another species. Which of the tools below is used to cut the gene from its normal location? A. Restriction enzyme B. Plasmid C. Bacteriophage D. Vector 3. Manufacturing recombinant DNA molecules involves cutting a gene from its normal location, inserting it into a circular piece of DNA from a bacterial cell, and then transferring the circle of DNA to cells of another species. Which of the below describe the circular piece of DNA from a bacterial cell? A. Restriction enzyme B. Plasmid C. Bacteriophage D. Vector

4. An cell that has the nucleus removed A. nucleated B. enucleated C. unucleated D. no such thing exists 5. The "donor nucleus" is taken from this type of cell Male Egg Somatic Enucleated 6. T or F Restriciton enzymes are used to cut DNA apart and help insert genes from one organisms into another 7.The 1st large mammal that was cloned Sheep Human Chimpanzee Frog

8. Restriction Enzymes will _____ DNA at ______ sequences paste, specific Cut, random Paste, random Cut, specific 9. T or F A commonon misconception is that if cloning humans ever happens, somone like Adolph Hitler could be cloned and the same person (personality and beliefs) would be brought back. Scientist have indicated that this would not happen 10.When genes are cut out, inserted and pasted, this is commonly known as Clonning Gel Electrophoresis Recombinant DNA Technology DNA Fingerprinting 11. Bacteria contian small circular segments of DNA called _______ that are very useful in genetic engineering Plasmids Vectors Restrction enzymes SNP’s

12. This is needed to help produce the clone, but it has no genetic connection to the clone somatic cell donor Surrogate mother Male sperm cell Morula 13. A “probe” in molecular biology is __________. a DNA or an RNA molecule used in hybridization reactions An instrumen t used to manipulate cells in culture A type of vector system A restrction enzyme

If we have an unknown segment of DNA that we wish to identify what technique would be best to determine this provided segment of nucleotides? DNA sequencing 2. If we would like to determine the genes expressed from a specific sequence what technique would be best? DNA microarray 3. When using recombinant DNA technology how can you be sure the cells you have isolated contain the recombinant plasmid you wish to work with? You can plate the cells on ampicillin agar plates. Only cells that have taken up the recombinant plasmid DNA will survive because they have the AMP resistant gene. The other cells do not and will die. 4. The creation of “Dolly” and other cloned animals having the exact same DNA are examples of what type of cloning? Reproductive cloning 5. In DNA sequencing what happens when a dideoxynucleotide is added to the strand being synthesized? How is this beneficial to the process? Synthesis stops. Strands of various lengths are created allowing the sequencing machine to indicate which nucleotide it stopped with and the length of this strand. The length helps us to determine where in the sequence the identified nucleotide is.