Laboratory techniques in molecular biology Flashcards
Primary culture
Prepared directly from tissues of an organismPrepared directly from tissues of an organism
Secondary culture
Sub cultured cells from primary culture
How bacteria produces large quantities of specific DNA or protein
To express protein inside recombinant gene into expression vector and then it’s introduced into the bacterium the advantage is that large amount of protein is easily
Gel electrophoresis
Technique used to separate DNA or RNA molecules of different sizes uses agarose gel DNA is loaded into precast Wells in an agarose gel which have been placed in an electrophoresis tank with buffer a current is then applied as the DNA is negatively charged at a neutral PH and it will migrate what’s the positive electrode through the gel DNA has a uniform charge to mass ratio and the fragments will separate according to size
The red of migration is determined by
Size of the DNA molecule concentration of the address there any confirmation strength of the electric field applied composition of the electrophoresis buffer
How do you make hand by seal
The use of etherium bromide for florist Florence under ultraviolet light when it is bound to DNA letters on the sides are the molecular weight markers with known sizes and can be used to estimate the sizes of the other bands
A DNA library is
A collection of cloned DNA fragments from a cell or tissue of an organism and can be created using different vectors which are generally kept in bacterial cells
Preparation of a DNA library
The DNA of the organism being used in the construction of the library is cut with a restriction enzyme at the same time the library plasmids are cut by the same restriction endonuclease the to cut some pills are then added together and the DNA is allowed to anneal DNA ligase is added to join the fragments and recombinant plasmids can then be introduced into bacterial cells the transformed bacteria are spread on all the plates containing antibiotic-resistant abiotic that the plasmids confer resistance to and therefore only the bacteria that have taken up placemats will grow rich separate bags of clothes should contain apartment of the different fragment of DNA from the organisms been used in the construction of the library different clothes grow as colonies on the plate in the bottom clones can be stored for later use
Difference between a genomic library and a cdna library
The genomic library is created from the genomic DNA of a cell line or tissue is cdna library is constructed from mrna and therefore only contains the protein-coding genes of the organism
Cloning of DNA
Synthesis of many copies of DNA
Plasmids are
Small circular DNA molecules that appear as single or multiple copies in bacterial cells
The DNA like is most widely used in the lab is
Bacteriophage T4
Pulse field electrophoresis
Similar to agarose gel electrophoresis separates even longer DNA molecules
