In a gene cloning project, the goal is to produce a recombinant plasmid DNA that

Question

In a gene cloning project, the goal is to produce a recombinant plasmid DNA that harbors the red segment of the genomic DNA specified in the following figure. The genomic Digesting this DNA with BamHI enzyme results in multiple cut fragments of 300 bp. 750 bp, 75 bp, and other fragments of unknown sizes at both tails. The plasmid DNA, on the other hand, is 2600 bp long and contains only one BamHI restriction site in the Tetracycline Resistant gene (TetR), such that insertion of any forign DNA in this site results in "insertional inactivation" of TetR. In a test tube we digest the genomic DNA and plasmid DNA with restriction enzyme Bar hope to bring the red segment (750 bp) into the BamHI cut site of the plasmid DNA. Then, frozen E. coli will be heat shocked in the presence of this plasmid such that the bacteria is transformed with plasmid DNA which we hope contains the red-insert segment. We then plate the bacteria solution on to three types of plates A) without any antibiotic, B) with both ampicillin and tetracycline antibiotic, and C) with ampicillin only DNA is 1600 bp, with multiple BamHI and EcoRI restriction sites mHI and then use the enzyme ligase with the 300 bp 750 bp 75 bp 420 bp 730 bp 450 bp BamHI EcoRI 2600 bp

Explanation / Answer

your queries are answered on sequential basis.

B) 75, 300, 750bp

c) both ampicillin and tetracycline

A) TetR is disrupted and ecoli cannot grow on tetracyclin.

QUERIES after this mention of a plate A,B and C whose imformation is not provided. please provide that and i may help u then.

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