batches of competent cells into electrocompetent cells for transformation. In the end, the competent cells would be utilized for both cloning purposes (to amplify your inserted gene ligated into pUC18). In the freezer at -80 C, you discovered numerous distinct Escherichia coli strains: BL21 competent cells One Shot BL21 Star (DE3) competent cells DH5a competent cells Rosetta™ host strains Which ones are appropriate for your work? Justify the decision you made.

batches of competent cells into electrocompetent cells for transformation. In the end, the competent cells would be utilized for both cloning purposes (to amplify your inserted gene ligated into pUC18). In the freezer at -80 C, you discovered numerous distinct Escherichia coli strains: BL21 competent cells One Shot BL21 Star (DE3) competent cells DH5a competent cells Rosetta™ host strains Which ones are appropriate for your work? Justify the decision you made.

Biology Today and Tomorrow without Physiology (MindTap Course List)

5th Edition

ISBN:9781305117396

Author:Cecie Starr, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Christine Evers, Lisa Starr

Chapter10: Biotechnology

Section: Chapter Questions

Problem 1CT

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Similar questions

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?
Ehat primer sets could be amplify the following DNA sequence? AATACGTCGCATGGggatccttttttatgcatg
Bacteriophage lambda is as one of the routinely used molecular cloning vectors, which alsoserved as a model system for the study of bacteriophage morphogenesis, DNA replication, andgene regulation.a) With the aid of a diagram, generally narrate how a foreign gene can be inserted into alambda insertion vector and subsequently infect an Escherichia coli cell.b) You are cloning a 7.5 kb gene into a lambda gt10 vector, utilizing a restriction site whichspecifically present in the vector. State the restriction site that you can use for this purposeand suggest a screening procedure to indicate successful integration of the gene into thehost genome.
You are engineering a new vector that contains a screenable marker that can be used for blue/white screening of successful clones. For each site (1, 2, and 3) on the cloning vector below, describe why it would or would not be a good place for you to put the polylinker to facilitate blue/white screening. You can assume that the polylinker itself will not interfere with coding sequence in that region. In other words, the polylinker length will be a multiple of 3 nucleotides, will not contain a stop codon, and any amino acids translated will not affect the activity of the protein in that region. The arrows indicate the direction of transcription for the gene.
You are engineering a new vector that contains a screenable marker that can be used for blue/white screening of successful clones. For each site (1, 2, and 3) on the cloning vector below, describe why it would or would not be a good place for you to put the polylinker to facilitate blue/white screening. You can assume that the polylinker itself will not interfere with coding sequence in that region. In other words, the polylinker length will be a multiple of 3 nucleotides, will not contain a stop codon, and any amino acids translated will not affect the activity of the protein in that region. The arrows indicate the direction of transcription for the gene. Note from student:As stated in the problem... "YOU CAN ASSUME THAT THE POLYLINKER ITSELF WILL NOT INTERFERE WITH CODING SEQUENCE IN THAT REGION. IN OTHER WORDS, THE POLYLINKER LENGTH WILL BE A MULTIPLE OF 3 NUCLEOTIDES, WILL NOT CONTAIN A STOP CODON, AND ANY AMINO ACIDS TRANSLATED WILL NOT AFFECT THE ACTIVITY OF THE PROTEIN IN THAT…
In preparation for transformation, why do you think we used calcium chloride and magnesium chloride in making the cells competent? Would your results be different if monovalent ions were used instead of divalent ions? Why do you think so?
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
1A) Which of the following best explains Griffith’s transformation experiments? Two strains of S. pneumoniae were used for the experiment. Griffith injected a mouse with heat-inactivated S strain (pathogenic) and living R strain (non-pathogenic). The mouse died and living S strain was recovered from the dead mouse. From this, it was later concluded that that mutation occurred in the DNA of the R-strain cells thus transforming them into a pathogenic S strain. Two strains of S. pneumoniae were used for the experiment. Griffith injected a mouse with heat-inactivated S strain (pathogenic) and living R strain (non-pathogenic). The mouse died and living S strain was recovered from the dead mouse. From this observation, it was later concluded that external DNA from the inactivated S strain was taken up by the R-strain cells thus transforming them into a pathogenic S strain. Two strains of S. pneumoniae were used for the experiment. Griffith injected a mouse…
You want to clone a 6,000 bp DNA fragment in E. coli. Which cloning vectors would be appropriate? How will you select transformants?
In bacterial transformation ... 1, Why do you have to incubate the competent bacterial cells on ice after the addition of plasmid DNA? 2. Why do you have to add SOC (Super Optimal Broth with Catabolite repression) medium to bacterial cells? 3. Why do you have to incubate the bacterial cells in SOC medium at 37oC for 45 minutes?
show your rationale How many RT-PCR products generated from 13 copies of mRNA (= cDNA) from 33 PCR amplification cycles? What was the original starting number of mRNA molecules if after 36 PCR cycles, the reaction had generated 996,432,412,672 copies?
The first publications describing the successful production and intracellular replication of recombinant DNA appeared in the early 1970’s. Since then, there is a long list of breakthroughs achieved by recombinant DNA technology. But which one of the following are NOT on that list? Select more than one answer ifneeded. a) Recombinant Factor VIII protein for the treatment of hemophiliacs b) Positional cloning of over 100 human disease genes c) Bacteria producing human insulin for the treatment of diabetic patients d) Reproductive cloning of over a dozen humans