You ligate a cDNA fragment with Bgl II and Xho I sticky ends into the Bgl Il and Xho I sites of the multi-cloning site within a vector (vector plus insert= 9 kbp). The vector has a resistance gene for kanamycin. Following the ligation, you transform the vector + insert (aka plasmid DNA) into chemically competent bacteria and spread the transformed bacteria on LB agar plates with antibiotic. The plates are incubated overnight at 37°C. If you spread the transformed bacteria on LB agar plates with ampicillin, what would you expect to see on the plate following a 24h incubation? Select one or more: J a. no bacteria/no growth O b. bacteria with vector + insert O c. insert alone d. bacteria with vector alone е. bacteria with no vector

You ligate a cDNA fragment with Bgl II and Xho I sticky ends into the Bgl Il and Xho I sites of the multi-cloning site within a vector (vector plus insert= 9 kbp). The vector has a resistance gene for kanamycin. Following the ligation, you transform the vector + insert (aka plasmid DNA) into chemically competent bacteria and spread the transformed bacteria on LB agar plates with antibiotic. The plates are incubated overnight at 37°C. If you spread the transformed bacteria on LB agar plates with ampicillin, what would you expect to see on the plate following a 24h incubation? Select one or more: J a. no bacteria/no growth O b. bacteria with vector + insert O c. insert alone d. bacteria with vector alone е. bacteria with no vector

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter13: An Introduction To Genetic Technology

Section: Chapter Questions

Problem 20QP: Analyzing Cloned Sequences A base change (A to T) is the mutational event that created the mutant...

See similar textbooks

Similar questions

For Pet41 (choose Pet41 a, b, or c as provided in the image) how would you design the primers (forward and reverse) for the following gene of interest and what restriction enzymes would be used (as shown in the image)? Be sure to explain and elaborate on why selected and how. Gene of Interest: atgggc gacaaaggga 241 cccgagtgtt caagaaggcc agtccaaatg gaaagctcac cgtctacctg ggaaagcggg 301 actttgtgga ccacatcgac ctcgtggacc ctgtggatgg tgtggtcctg gtggatcctg 361 agtatctcaa agagcggaga gtctatgtga cgctgacctg cgccttccgc tatggccggg 421 aggacctgga tgtcctgggc ctgacctttc gcaaggacct gtttgtggcc aacgtacagt 481 cgttcccacc ggcccccgag gacaagaagc ccctgacgcg gctgcaggaa cgcctcatca 541 agaagctggg cgagcacgct taccctttca cctttgagat ccctccaaac cttccatgtt 601 ctgtgacact gcagccgggg cccgaagaca cggggaaggc ttgcggtgtg gactatgaag 661 tcaaagcctt ctgcgcggag aatttggagg agaagatcca caagcggaat tctgtgcgtc 721 tggtcatccg gaaggttcag tatgccccag agaggcctgg cccccagccc acagccgaga 781 ccaccaggca gttcctcatg tcggacaagc ccttgcacct…
You have isolated a cDNA clone encoding a protein of interest in a higher eukaryote. This cDNA clone is not cleaved by restriction endonuclease EcoRI. When this cDNA is used as a radioactive probe for blot hybridization analysis of EcoRI-digested genomic DNA, three radioactive bands are seen on the resulting Southern blot. Does this result indicate that the genome of the eukaryote in question contains three copies of the gene encoding the protein of interest? Explain.
Once cloning is complete, a recombinant OXA-M290 protein can be produced in E. coli. Competent E. coli BL21(DE3) cells are transformed with the pET-28a-OXA-M290 plasmid, and the transformed cells are grown on an agar plate that contains kanamycin. The E. coli BL21(DE3) cells transformed with the pET-28a-OXA-M290 vector are next grown in liquid growth media that contains kanamycin. Once the culture reaches an OD600 of 0.6, isopropyl β-D-1-thiogalactopyranoside (IPTG) is added to a final concentration of 0.5 mM to induce protein production. How does the addition of IPTG result in the transcription of genes that are regulated by the T7 promoter in E. coli BL21(DE3) cells? [4 – 6 sentences suggested] For Context ONLY: For this biology course and based on the results obtained from a previous BLAST search, the bacteria responsible for the infection at a Hospital is most likely a new strain of Escherichia coli. This strain is given the name E. coli AFB. Earlier, BLAST was used to study a…
What is the minimum and maximum cDNA input from FFPE samples in library preparation phase to perform the RNA SEQ by Ion Proton?
When using a conventional plasmid cloning vector containing a b-galactosidase gene, it is possible to perform a "blue-white screen" to determine which bacteria have taken up a plasmid into which a DNA fragment as been inserted, as opposed to those that have taken up just reclosed plasmid vector, by growing the transformed cells on nutrient agar plates containing the artificial b-gal substrate X-gal. Will bacteria that have taken up a plasmid into which a DNA fragment has been inserted form a blue colony or a white colony when grown on this medium? Briefly explain why these bacteria would form a colony of the color you chose.
Analyzing Cloned Sequences A base change (A to T) is the mutational event that created the mutant sickle cell anemia allele of beta globin. This mutation destroys an MstII restriction site normally present in the beta globin gene. This difference between the normal allele and the mutant allele can be detected with Southern blotting. Using a labeled beta globin gene as a probe, what differences would you expect to see for a Southern blot of the normal beta globin gene and the mutant sickle cell gene?
Your cloning vector has restriction recognition sites for two restriction endonucleases, EcorI and BamHI. However, the DNA to be manipulated does not have recognition sites for these two restriction endonucleases. How would you construct a recombinant DNA for the given DNA?
You have ligated a foreign DNA fragment into the EcoRI site in multiple cloning site of pUCI18. You then transform how E.coli with the ligation mixture. Some of the bacterial colonies growing on the nutrient agar plate that contains ampicillin and Xgal are white and some are blue. Bacterial colonies that are white a) contain recombinant plasmid DNA b) have hydrolyzed ampicillin to form a colored compound c) are untransformed bacteria d) contain non recombinant plasmid DNA
Explain the process of how X-gal screening works with pUC19, you may build a model with boxes and arrows. 2. You are utilizing BamHI (GGATCC) restriction site and HindIII (AAGCTT) restriction site. Within pUC19, BamHI is at position 263, while HindIII is at position 233. (Hint: position is like coordinate on a map). If you manage to insert GTF2H5 in pUC19 vector, what are the sizes of fragments if you digest the pUC19-GTF2H5 (this is after insertion) with the following restriction enzyme combination after gel electrophoresis: BamHI and HindIII There is an NdeI site right in the middle of the GTF2H5 that has been inserted in pUC19, what would be the fragment sizes, if you digest with BamHI and NdeI. HindIII and NdeI BamHI, HindIII, and NdeI. 3. Design an experiment to confirm the presence of insert GTF2H5 in pUC19 vector using the following method (besides restriction analysis above), assuming that you know the sequence of GTF2H5: Southern Blot Polymerase Chain Reaction
Early gene-cloning experiments involved insertion at one restriction site in the vector; for example, the insert would have an EcoRI site at each end, and the vector would be opened at an EcoRI site prior to ligation. Under what circumstances would asymmetric cloning be desirable, with the insert having a different restriction site at each end
Which restriction enzyme is best suited for cloning in pUC8?
What would three possible reasons be that a PCR wouldn't work after cloning a TAQ sequence into a His-tag vector making three different constructs and verifying via SDS-PAGE that the TAQ protein is there?