Mariah+Pierce

= Part 1: DNA cloning/ Recombinatn DNA/ Genetic Engineering = || ||  ||
 * Procedure || Gene Cloning with Bacterial Plasmids || Nucleic Acid Hybridization || Genomic Library || Polymerase Chain Reaction (PCR) ||
 * Description || * DNA is cloned in a lab
 * Done //in Vitro//
 * Foreign DNA is inserted
 * Gene Cloning is the production of multiple copies of a single gene || * Is the screening of a library for clones carrying a Gene of interest
 * probes are labeled w/ a radioactive isotope
 * stored in multiwell plates
 * Cells are transferred on a membrane made of nylon or nitrocellulose
 * Once desired gene is located, we can grow identical cells in a liquid culture || * Is the storing of cloned gene in DNA libraries
 * Stored as a collection of phage clones
 * Stored in multiwelled plastic plates- one clone per well
 * Made //in vitro// || * Made //in vitro//
 * Specific segments can be quickly amplified in a test tube
 * "Is like photocopying just one page rather than checking out every single book in a library" -Campbell AP Bio Text
 * With each recessive cycle the number of target segment molecules of the correct length doubles soon surpassing all other DNA molecules ||
 * Analyze || * Plasmid isolated from bacterial cell
 * Foreign DNA is inserted into plasmid
 * The resulting plasmid then has recombinant DNA (DNA from two sources)
 * The plasmid is then returned to a bacteria cell
 * This cell is known as a recombinant bacterium
 * Recombinant cell reproduces through repeated cell division and makes clones
 * Foreign DNA and any genes it carries are cloned at the same time
 * Restriction enzymes are needed
 * Enzymes that cut DNA molecules at a limited number of specific locations
 * Methyl groups (-CH3) can be added to adenines of cytosines
 * Restriction sites are also present
 * Sequences of nucleotides is same on both strands when read from the 5' -> 3' direction
 * There is also a sugar phosphate backbone that is cleaved by the restriction enzymes || * Nucleic Acid Hybridization's main function is to detect a genes DAN by its ability to base-pair with a complementary sequence on another nucleic acid molecule
 * Nucleic acid probes are present
 * the complementary molecule that is known, a short single stranded nucleic acid that can either be DNA or RNA
 * Another function: If we know at least one part of the nucleotide sequence of teh gene of interest, we can synthesize a probe complementary to it
 * Example: part of gene of interest 5'...GGCTAACTTAGC...3', so the probe would be 3'...CCGATTGAATCG...5'
 * In other words, each probe molecule will hydrogen bond to the complementary sequence (desired gene) || * Cloning procedure- "Shotgun approach"
 * Starts with a mixture of fragments from entire genome of an organism
 * No single gene is targeted for cloning
 * "Plasmid clones" each thought of as a book in the library containing specific info
 * Bacteriophages are also involved and have also been used as cloning vectors for making genomic libraries
 * Restriction enzymes and DNA ligase are also a part of the genomic library; they trim down fragments of foreign DNA
 * Another vector type is the Bacterial Artificial Chromosome: are simply large plasmids
 * Capable of carrying inserts of about 100-300kb
 * Another type of library is the cDNA (Complementary DNA) library
 * Starts with mRNA extracted from cells
 * Uses the enzyme: Reverse transcriptase to make single stranded DAN transcriptase out of mRNA molecules
 * Another part of the cDNA library in the Poly-A tail of the 3' end
 * It allows the use of the short strand of dT (thymine deoxyribonucleic acid) as a primer for the reverse transcriptase
 * DNA polymerase is also a part of this library; it synthesizes a DNA strand complementary to the first || * Specific target segments are an important part, because they are what becomes amplified during PCR
 * Automation helps PCR make billions of copies of a target segment of DNA in a few hours
 * There are three main parts to each cycle that help produce and exponentially growing population of identical DNA molecules
 * The reaction mixture is heated to denature
 * Then it's cooled to allow annealing (hydrogen bonding)
 * Finally, a heat stable DNA polymerase extends the primers in the 5' -> 3' direction
 * A key component to PCR would be the minute amounts of DNA needed to start the process
 * Another key element would be complete target sequences
 * Primers are also important in PCR specificity
 * Must be 15 or so nucleotides long ||
 * Apply- What can you do with it, and how can it be used? || * Gene cloning is the production of multiple copies of a single gene to produce a protein product
 * Isolated copies can be cloned for specific uses such as basic research
 * Endowing an organism with a new metabolic capability such as pest resistance in crops
 * You can alter bacteria for cleaning up toxic waste
 * you can create more proteins that dissolve clots in heart attack therapy
 * You can treat stunted growth || * We can screen a large number of clones simultaneously for the presence of DNA complementary to our DNA probe
 * Discover the location of a clone carrying the desired gene
 * Use the cloned gene as a probe to identify similar or identical genes in DNA from other sources, such as another bird species || * You can clone a gene without knowing what the cell type expresses or if you are not able to obtain enough cells of the appropriate type
 * This process is perfect if the sequences are absent from the fully processeed mRNAs used in makindg a cDNA library || * PCR is a quicker and more selective process of DNA cloning
 * It is best used when DNA is scanty or impure
 * With automation, PCR can make billions of copies of a target segment of DNA within a few hours
 * PCR is being used increasingly over the years
 * You can make enough of a specific DNA fragment to insert directly into a vector
 * May entirely skip the making and screening of a library
 * ;Produces an exponentially growing population of identical DNA molecules
 * Used to amplify DNA from a wide variety of sources
 * DNA from fingerprints
 * Tiny amounts of blood
 * Tissues
 * Semen from crime scenes
 * Cells for parental diagnosis of genetic disorders
 * Defected viruses: HIV ||
 * Synthesize- What does it make you think of? || * This Procedure is similar to that of a blood transfusion or a bone marrow transplant in the fact that each endow an organism with a new metabolic capability || * Crime scene DNA screenings
 * Similar to the process of using a magnet to find metal in the way both are attreacted to the other (metal to magnet, nucleotides of gene of interest to nucleotides of probe) || * DNA screening
 * Genetic Counseling || * Crime Scene investigations
 * Genetic Counseling ||
 * Argue || This procedure is very important because it helps many people perfect certain tasks such as creating pest resistant crops. It has also helped to contol stunted growth and the production of needed proteins. || Thsi process is important because we can use the clone gene as a probe to identify similar or identical genes in DNA from other sources. || This process is important becasue it can help to find the gene of interset using probes. This process can help couples with genetic counseling, or the screening of families with genetic disorders in their genes in order to recieve advice on consequences of child conception. || The DNA cloning procedure PCR is important because, again, it helps couples with genetic counseling. It is also important because it can be used in forensics for cime scenes. ||
 * Picture || [[image:riedell-dna-tech-project-2012/genomic.jpg width="299" height="663"]] || [[image:riedell-dna-tech-project-2012/NUCLEIC.gif width="346" height="383" align="center"]]
 * Source || [] || h[|ttp://www.accessexcellence.org/RC/VL/GG/nucleic.php] || [] || [] ||