can you please answere the questions above. they are the questions that I need t

Question

can you please answere the questions above. they are the questions that I need to answer for my transformation experiment which was this:

Exogenous DNA does not pasoively enter E coli cells thunt are not competent. What treatment cells require to be compwarnt? 2. The recovery broth used in his experiment did not contain ampicillin. Why? 3. The antibiotics kanamydn and betracydine, interfern with trans- lation by binding to the ribosoenas. Competent E. coli cens canbe transformed with plasmids enooding resistance to these drugs. When the censzre plated on agar media containing kanamycinar tetracycline, salellite amionies ene not fourd. Increasing incuba- tion times anabe amount of odisplatek do not give ise satel lites in this case. Why?

Explanation / Answer

TRANSFORMATION EXPERIMENT :

A) Preparation of competent cells : The host bacteria i.e., Ecoli should make to intake of the recombinant plasmid, inorder to facilitate the bacteria cell is changed to the Competent cell which means, removal of cell wall so that the DNA facilitate into the cell, for the competent cells are prepared as follows:

B) The host cell is sensitive for antibiotics, so these are taken with normal medium not under antibiotic selective pressure.

C) When the plasmid containing Kan R genes and Tet R genes confer into the competent cell, this bacteria will be resistant for that antibiotic selective pressure, hence the medium is provided with selective antibiotic pressure, so that recombinant cells and non recombinants are identified easly, non recombinants dont have plasmid so they cannot survive under selective antibiotic pressure. Which shown in above your diagram.

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