2. The plasmid cloning vector Pbr322 (please refer to the figure in the lecture

Question

2. The plasmid cloning vector Pbr322 (please refer to the figure in the lecture slide) is cleaved with the restriction endonuclease Pstl. The cloned DNA fragment cut by Pstl is 1,000 bp long and has an EcoRI site 250 bp from one end. Two different recombinant plasmids are cleaved with EcoRI and analyzed by gel electrophoresis (Lane 1 and 2), giving the patterns shown below. Note that in pBR322, the Pstl and EcoRI restriction sites are about 750bp apart. The entire plasmid with no cloned insert has 4,361 bp The size markers in lane 4 have the number of nucleotides noted on the right. Determine the correct size of nucleotide in DNA fragments A, B, and C. (15 pt) 4length 3,000 1,000 750 s00 250

Explanation / Answer

The total size of the plasmid with no cloned insert is 4361 bps.

The size of DNA fragments A is 4000 bps and size of C is 1361 bps

The size of DNA fragment B is 4361 bps while the other band in the same lane is 1000 bps.

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