please write in detail the lab procedure of how to see the e

please write in detail the lab procedure of how to see the expression of pglo in eukaryotic cells by TRANSFECTION. please do not give transformation, i need transfection.

Solution

Transfeciton process for expression pglo plasmid: The pGLO plasmid has GFP gene which expresses Green fluorescent protein. This protein is expressed when the plasmid gets a host. We can transfect the whole plasmid without using bacteria or virus by various methods. We can use chemical method like calcium phosphate or Lipofection method. Lipofection involves use of liposome. These chemicals easily absorbed through the membrane. The DNA has negative charge so calcium binds with it and liposome has structural similarly with plasma membrane. The other method can use such as electroporation which involve use of electric current for some time to allow the looseness of membrane and then readily available to take the DNA inside. Another method sonoporation is also used in which ultrasound is used to lose the membrane for allowing insertion. The transfeciton process is quick method to achieve, electroporation sonoporation and chemical methods generally tool 10-15 seconds to make the process complete.