http://blog.addgene.org/plasmid-101-origin-of-replication http://dnasu.org/DNASU

Question

http://blog.addgene.org/plasmid-101-origin-of-replication

http://dnasu.org/DNASU/GetVectorDetail.do?vectorid=60

Once you ligate (or piece together) the happy gene into the vector you then place it into a host cell (for this experiment, E. coli). How would you know which bacterial cells contain the vector? The process of transformation (placing genetic material into a host cell) results in only a fraction of the r containing the vector. 9. (3 points) Describe whattype of media you would plate yourtransformed cells in order to allow for only the cells containing your happy-gene-containing-vector to grow. List the components of your media and why you chose them.

Explanation / Answer

use a plasmid whic has a selectable marker gene in the MCS which would get disrupted n insertion of the happy gene therby allowing you to screen against the cells possessing this selectable marker.

9) glucose = carbon source

peptone = protein source

beef extract = salts, vitamins, carbohydrates and nitrogen

NaCl = for isotonicity

Distill water = to dissolve all the media component

ampicillin (selectable marker - ampicilin resistance - gene outside MCS) = will help to select transformants but will not gaurantee that the tranformants have the recombinant plasmid or not

agar = solidifying agent

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