last of genetics

• Recombinant DNA: molecules that are produced in a test tube by covalently linking DNA fragments from two different sources.
• Vector: a small DNA molecule that is used as a carrier of a DNA segment in a cloning experiment.
• Host Cell: a cell that is infected with a virus or bacterium, or one that harbors a vector.
• Plasmid: a general term for a DNA molecule (most often circular) that exists independently of the chromosomal DNA. Some plasmids are used as vectors in cloning experiments.
• Selectable Marker 


  a gene that provides a selectable phenotype in a cloning experiment. Many selectable markers are genes that confer antibiotic resistance.
• Restriction Enzyme 


  an endonuclease that cleaves DNA. The restriction enzymes used in cloning experiments bind to specific base sequences and then cleave the DNA backbone at two defined locations, one in each strand. Also referred to as a restriction endonuclease.
• Sticky End 


  DNA fragments that hydrogen bond to each other due to their complementary sequences and are created by most restriction enzymes
• Blunt ends 


  the opposite of sticky ends
• DNA Ligase: Enzyme that covalently bonds the DNA backbone together
• Competent Cells


  cells that can take up DNA from the environment or an extracellular medium.
• reverse transcriptase 


  an enzyme that uses an RNA template to make a double-stranded DNA molecule
• oligonucleotide: a short strand of DNA, typically a few or a few dozen nucleotides in length.
• cDNA: complimentary DNA that is made from an mRNA template by the action of reverse transcriptase.
• Genomic Library


  a DNA library of recombinant vectors that carry chromosomal DNA fragments.
• cDNA library


  a DNA library whose recombinant vectors carry cDNA inserts.
• Taq polymerase


  a thermostable form of DNA polymerase used in PCR experiments.
• Primers


  a short segment of DNA or RNA that initiates DNA replication.
• Annealing


  in PCR, the process in which a primer binds to a strand of template DNA.
• Thermocycler


  a device that automates the timing of temperature changes in each cycle of a PCR experiment
• PCR


  the method for amplifying a DNA region involving the sequential use of oligonucleotide primers and Taq polymerase.

  DNA is denatured at high temps, then the primer is annealed and finally, the primer is extended to allow for DNA synthesis
• TaqMan 


  specific probe used in quantitative PCR, requires a TaqMan primer. Has a fluorescent reporter molecule at one end (Reporter) and a molecule that blocks the fluorescence (Quencher). Fluorescence increases in proportion to the amount of PCR product produced.
• Real-time PCR


  a method of PCR in which the synthesis of DNA is monitored in real time. This method can quantitate the starting amount of DNA in a sample
• Site-directed mutagenesis 


  a gene-editing technique that allows a researcher to produce a mutation at a specific location within a cloned DNA segment by using an oligonucleotide primer with a mismatch.
• Antibody


  proteins that are produced by the immune systems of vertebrates, which recognize foreign material (namely, viruses, bacteria, and so forth) and target it for destruction; also known as immunoglobins.
• Dideoxy DNA Sequencing


  a method of DNA sequencing that uses dideoxyribonucleotides to terminate the growth of DNA strands.
• Reverse transcriptase PCR


  a modification of PCR in which the first round of replication involves the use of RNA and reverse transcriptase to make a complementary strand of DNA.
• Northern blotting 


  a technique used to detect a specific RNA within a mixture of many RNA molecules.

  Gel: RNA, Probe: DNA or RNA
• Western Blotting


  A technique used to detect a specific protein within a mixture of proteins
• Western Blotting


  • Can determine if a specific protein is made in a particular cell type
  • Can determine if a specific protein is made at a particular stage of development
• Primary antibody


  Recognizes the protein of interest
• Secondary antibody


  Conjugated to alkaline phosphatase, recognizes the constant region of the primary antibody
• Western Blotting

  1. Protein in gel

  2. Primary antibody binds protein

  3. Secondary antibody binds primary antibody

  4. Alkaline phosphatase converts colorless dye to black compound

  5. Proteins of interest indicated by dark bands
• Transformation


  When the cell takes up the plasmid
• CRISPR-Cas


  a system found in bacteria and archaea that provides defense against viruses, plasmids, and transposable elements. Can be used to introduce mutations into genes.
• GMO


  an organism that has received genetic material via recombinant DNA technology.
• Transgenic Organism 


  an organism that has DNA from another species incorporated into its genome via recombinant DNA techniques.
• Biological Control


  the use of microorganisms or products from microorganisms to alleviate plant diseases or damage from environmental conditions (e.g., frost damage).
• Bioremediation


  the use of living organisms or their products to decrease pollutants in the environment.
• Gene Replacement


  the swapping of a cloned gene made experimentally with a normal chromosomal gene found in a living cell.
• Gene Knockout


  in the case of diploid species, the condition in which both copies of a gene have been altered to an inactive form.