Course Guide Chapter 18: Recombinant DNA Technology Flashcards
What is the function of restriction endonucleases?
enzymes located from prokaryotes that recognize a specific DNA sequence and cleave the DNA at that recognition site or another place
What is the function of DNA polymerase?
used to make DNA in vitro
What is the function of DNA ligase?
catalyze the covalent bonding of the 3’ and 5’ ends of two DNA strands
What is the function of reverse transcriptase?
an RNA dependent DNA polymerase; uses an RNA template to make a complementary DNA
What is the purpose of gel electrophoresis?
used to fractionate DNA or RNA fragments based on their size
What is the purpose of restriction fragment length polymorphism?
used to identify differences in the restriction pattern of a specific gene or DNA region between several individuals of a species or several related species
What is the purpose of radiolabeling DNA fragments?
using a radioisotope or fluorescent label to identify a DNA fragment in an organism
What is the purpose of DNA sequencing?
an in vitro DNA synthesis using a DNA template is done to extend the new strand of DNA from the primer in a suitable buffer
What is the purpose of polymerase chain reaction (PCR)?
used to amplify large amounts of DNA from small amounts of samples
What is southern hybridization?
hybridizing a labeled DNA probe to the target DNA sequences immobilized on a nylon membrane; used to study the genome organization of a particular gene and determine the approximate number of copies of a specific gene
What is northern hybridization?
hybridizing a labeled DNA probe with size-fractioned RNA immobilized on a nylon membrane; used to determine the RNA levels for a particular gene at different stages or in different tissues or organs of an organism
What is western hybridization?
involves protein-antibody interaction; a specific protein (antigen) is recognized first by a primary antibody which is then recognized by a secondary antibody conjugated to a fluorescent probe or an enzyme. This enzyme can convert a chromogenic substrate into a colored substance for the detection
