In cloning a specific fragment from a mixture of different fragments of DNA, three classes of plasmids can be produced: vector containing the desired fragment (gene of interest), vector containing other fragments, and re- ligated vector containing no inserted DNA. What class of vector would you expect to find at the highest frequency? Multiple Choice O All three types of vector will be found in approximately equal proportions Vector containing random genomic DNA fragments Vector containing the gene of interest Vector with no insert

Below is a plasmid with restriction sites for Baml and EcoRI, Several restriction digests were done using these two enzymes either alone or in combination. Hint: Begin by determining the number and size of the fragments produced with each enzyme. "kb" stands for kilobases, or thousands of base pairs. Plasmid Gel lanes I | III IV V 6 Kb Bam HI Bam HI. 20 Kb PGEN101 (20 Kb) 2 Kb 11 Kb Bam HI 8 Kb 8 Kb 4 Kb 6 Kb Eco RI 3 Kb 21. Which lane shows a digest with BamHl only? a. I b.I c. II d. IV e. V 22. Which lane shows a digest with EcoRLonly? a. I 23. Which lane shows the fragments produced when the plasmid was incubated with both EcoRI and BamH1? a. I b.I c. II d. IV e. V b. I c. II d. IV e. V Base pairs

Clone number in this case is number 196 as shown in the images. What is the exact length of the segment of human DNA that has been inserted into the plasmid? *report the entire length of the insert, not just the sequences matching the ends and labels of wells isn't needed for answer*

Primers designing for epitope tagging: Design forward and reverse primers to amplify the following gene with 6×HIS-tag on the N-terminus of the protein. To be cleaved and inserted into the plasmid, add restriction sites for EcoRI and HindIII at 5' and 3'. ATGCTCTCCGCCCTCGCCCGGCCTGTCAGCGCTGCTCTCCGCCGCAGCTTCAGCACCTCAGCC CAGAACAATGCTAAAGTAGCTGTGCTAGGGGCCTCTGGAGGCATCGGGCAGCCACTTTCAC TTCTCCTGAAGAACAGCCCCTTGGTGAGCCGCCTGACCCTCTATGATATCGCGCACACACCC GGAGTGGCCGCAGATCTGAGCCACATCGAGACCAAAGCCGCTGTGAAAGGCTACCTCGGAC CTGAACAGCTGCCTGACTGCCTGAAAGGTTGTGATGTGTAA

The plasmid cloning vector pBR322 is cleaved with the restriction endonuclease PstI. An isolated DNA fragment from a eukaryotic genome (also produced by PstI cleavage) is added to the prepared vector and ligated. The mixture of ligated DNAs is then used to transform bacteria, and plasmid-containing bacteria are selected by growth in the presence of tetracycline. The cloned DNA fragment is 1,000 bp long and has an EcoRI site 250 bp from one end. Three different recombinant plasmids are cleaved with EcoRI and analyzed by gel electrophoresis, giving the patterns shown below.   What does each pattern say about the cloned DNA?   Note: pBR322, the PstI and EcoRI restriction sites are about 750 bp apart. The entire plasmid with no cloned insert is 4,361 bp. Size markers in lane 4 have the number of nucleotides noted.

Clone number in this case is number 196 which is shown in the images.  State whether a BamHI site has been re-created at the forward- and the reverse-end junctions of the human DNA with the plasmid vector  band sizes are shown in one of the images. (0.5, 1, 2, 3, 4, 5, 6, 8, 10kb)

An optimum ligation reaction should contain approximately 50 ng of vector. Given yourconcentrations recorded below, what is the volume of vector that should be added to eachligation reaction to have this mass of DNA in the reaction? Size of asPink-promoterless in bp: 702 (vector) Size of pCusC in bp: 157 The concentration of digested & purified PCR insert: 13.849 The concentration of digested and purified plasmid: 6.887 Any help with this is appreciated. I'm a bit confused thx

The restricition EcoRI cleaves double-stranded DNA at the sequence 5'-GAATTC-3' the restriction enzyme Hindll cleaves at the sequence 5'-AAGCTT-3'. A 20 kb circular plasmid containing OFP gene is digested with each enzyme individually and then in combination, and the resulting fragment sizes are determined using electrophoresis Restriction Enzyme(s) ECORI alone Fragment length 6 kb ,14 kb 7 kb , 13 kb 2kb, 4kb, 5 kb, 9kb Hindlll alone EcoRI and Hindll Construct the circular molecule and indicate the relative positions of the EcoRI and Hindlll restriction sites

Do virtual restriction digests of 300ng pGLO plasmid using HinDIII, EcoRI, and XhoI separately, and a double digest of pGLO with HinDIII plus EcoRI.  How many nanograms of DNA should we expect to be contained within the slowest bandfrom the HinDIII-digest of pGLO?

Please both parts