Lab 7 Flashcards
Traditional biotechnology
Has been utilized for centuries and includes selective plant and animal breeding and the use of yeast in fermentation to produce cheese, bread and beer and wine
Modern biotechnology
Has been developing since the 1970’s and specifically refers to the manipulation of DNA in vitro ( in glass, test tube or dish) modern techniques permit the alternation of specific DNA sequences as well as transfer genes between organisms
Nucleotide
One of the structural components or building blocks of DNA and RNA
Each nucleotide is composed of three parts:
-a nitrogenous base ( one of adenine, cytosine, guanine or thymine )
-a sugar (deoxyribose)
-a phosphate group, which is negatively charged
Nitrogenous base
Adenine, cytosine, guanine or thymine
Deoxyribose
Sugar
Hydrogen bond
Attraction between hydrogen atoms
Complementary
A property of double-stranded nucleic acids such as DNA and RNA as wells as DNA-RNA duplexes
Central dogma
The information that is encoded in the DNA is transcribed into a complementary RNA copy. The RNA copy is translated into a sequence of amino acids, which are building blocks of protein. This flow of information from DNA to RNA to protein is called the central dogma.
DNA - RNA- (Amino acids) protein
Nucleated
Cells with nucleus
Isotonic
When two solutions have the same solute concentration
Saline
A solution of salt and water
Polymerase chain reaction PCR
Technique used to rapidly make many copies of a DNA molecule or part of a NDA molecule
Four ingredients required for Polymerase chain reaction PCR
- DNA extract
- Each of the four deoxyribonucleic triphosphates (dNTPs: dATPs, dCTPs, dGTPs, dTTPs) these are building blocks used to produce copies of DNA
- Primers- short segments of synthetic DNA necessary for the initiation of DNA replication
- DNA polymerase (Taq polymerase) - a heat stable enzyme that elongates the DNA chain by adding dNTPs to the primers
The ingredients are combined and then placed in a thermal cycle, an automated system that maintains a series of temperatures for specific time periods
Denaturing DNA (PCR cycle step 1)
Heat to separate the two strands of the DNA double helix
Anneal
The process of heating and cooling two single stranded oligonucleotides with complementary sequences
Deoxyribonucleotide triphosphate dNTP
dATP
dCTP
dGTP
dTTP
DNA/Taq polymerase
A heat stable enzyme that elongates the DNA chain by adding dNTPs to the primers
Thermal cycler
An automated system that maintains a series of temperatures fro specific time periods
Extraction
DNA getting extracted from nuclei of cells
Restriction fragment analysis
Comparison of nucleotide sequences of different amplified DNA sequences, using several restriction enzymes that “cut” the DNA at different recognition sites
Recognition sequence/restriction site
Specific site where restriction enzyme cuts dna
Restriction digest
The first procedure used in restriction fragment analysis
Where a restriction enzyme is added to the PCR product and the solution is placed in an incubator, the enzyme cuts the DNA in the PCR product into specific fragment sizes and number, depending on number pf restriction sites present
Restriction enzyme
EcoR1
An enzyme that recognizes specific DNA sequences and cleaves the DNA molecule at this recognition site
Gel electrophoresis
Technique for separating macromolecules (eg fragments of DNA) using an electrical current and a porous matrix, based on speed of migration of the molecules which is mainly influence by their size
Agarose
Substances used in gel for electrophoresis
Ethidium bromide
Sometimes added to running buffer during the separation of DNA fragments by agarose gel electrophoresis
Buffer solution
Water solvent based solution
DNA fingerprint
DNA fingerprinting is a laboratory technique used to determine the probable identity of a person based on the nucleotide sequences of certain regions of human DNA that are unique to individuals
Ladder
Contains DNA fragments of known size
Paternity test
The same DNA analysis described above for criminal cases can be used for paternity tests
DNA from a child that is cut by a restriction enzyme will result in restriction fragments that are either their mothers or their fathers restriction fragments
Recombinant DNA
DNA molecule carrying genes derived from more two or more sources (two species)
Genetically modified organism GMO
A microbe whose DNA has been altered using genetic engineering techniques; recombinant DNA
Transgenic
Refers to an organism or cell whose genome has been altered by the introduction of one or more foreign DNA sequences from another species by artificial means
Bacterial transformation
A process of horizontal gene transfer by which some bacteria take up foreign genetic material (naked DNA) from the environment
Plasmid
A small circular DNA molecule found in bacteria and some microscopic organism
Green fluorescent protein GFP
Fluorescent makers of gene expression and for determination of protein and motility in living cells
Annealing of primers (PCR cycle step 2)
Cool so that primers can bond to the single strand of DNA
Extension of primers (PCR cycle step 3)
Heat to allow DNA polymerase (Taq) to add dNTP’s to then of the primers
