The Mechanism of Transformation with Competent Cells

transformation is the process by which DNA is introduced into bacteria bacteria can then replicate and express that DNA which can be isolated for downstream applications artificial transformation of bacteria involves producing competent cells capable of exogenous tiene uptake there are two primary methods of artificial transformation used in the laboratory chemical transformation and electroporation it is thought that chemical transformation which requires chemically competent cells uses divalent cations to increase the permeability of the bacterium cell wall thereby increasing the likelihood of DNA acquisition many common protocols include a heat shock step to improve DNA uptake electroporation involves the application of an electrical field to the competent cells in suspension thereby introducing holes in the cell's perimeter the newly compromised cell wall allows exogenously na to pass into the cytosol of the bacteria the bacteria following transformation allows for repair and recovery of the cells in liquid medium without antibiotic after outgrowth the culture is spread on selective plates to isolate transform ins containing the newly acquired DNA which conveys antibiotic resistance a variety of competent ecoli types are available in different formats and for different applications such as high efficiency cloning and protein expression visit clone with any be com for the full list of products available for this application

Channel: New England Biolabs

Views: 262,557

Keywords: transformation, cloning, competent cells

Id: 7Ul9RVYG5CM

Length: 1min 42sec (102 seconds)

Published: Fri Jun 13 2014