University:
Western Oregon University
Course:
BI 102 | General Biology
Academic year:

2022
Views:

10

Pages:

1
Author:

LauraParks11

Applications of PCR: PCR: - - technique for the in vitro amplification of specific DNA sequences -can make billions of copies of the target sequence of DNA in a few hours - it is a cycle of three steps of heating, cooling, and replication - cycle is repeated many times to produce multiple copies of the original DNA cloning a desired gene amplifying cloned DNA from a vector creating mutations in clone genes amplifying old DNA detecting bacterial or viral infection genetic diagnosis forensic science and paternity testing Components: Target dsDNA/ 2 oligonucleotide primers/ thermostable DNA polymerase/ DNTP/ reaction buffer The 3 PCR steps: 1- DNA denaturation 2- primers annealing 3- DNA polymerase (Tag) extends the DNA chain => repeat the cycle 25-35 times D 2) A Process might be affected from contamination PCR products are then analyzed by Gel Electrophoresis A RB Gel electrophoresis: is a technique for visualization and analysis of DNA where DNA fragments are separated into distinct bands by size in a gel matrix (agarose gel made from seaweed) by applying an electrical field [seen because of chemicals like ethidium bromide]; the DNA move from the negative to the positive side, and the smaller fragments are faster than the larger ones 4) Sample preparation and loading: mix the samples of the DNA with the 6x loading dye which allows the samples to be seen and it increases the density causing the DNA to sink into the gel walls Method: 1- prepare agarose gel 2- submerge agarose gel in gel tank containing buffer that conducts electricity 3- mix the DNA sample with loading dye 4- load the DNA sample into the wells 5- apply electrical charge 6- place gel on trans illuminator 7- photograph gel for analysis they - + will ask for - t - - t t size . - t - t - t t