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2. Mutation frequency A culture of E. coli is going to be mutagenized with transposons for...
A. You have been given a tube of E. coli. You are asked to make 1...
A. You have been given a tube of E. coli. You are asked to make 1 mL total volume of 10-1 dilution of the bacterial culture. Explain how you would do this. Show all necessary calculations. ____ ml cells + _____ ml water = 1 ml (total volume) B. Next, you were asked to make a 10-2 dilution of the bacterial sample. Explain how you would perform this. Show all necessary calculations. You have bacteria at a concentration of 2...
6. Transposon mutagenesis was performed to identify genes that are involved in the biosynthesis of theA amino acid alanine. A wildtype culture of E. coli was infected with a defective phage carrying...
6. Transposon mutagenesis was performed to identify genes that are involved in the biosynthesis of theA amino acid alanine. A wildtype culture of E. coli was infected with a defective phage carrying Tn10 A and the culture was plated on LB-tet media to obtain tet-resistant colony forming units. (LB is a rich growth media for E. coli and contains all amino acids required for bacterial growth) Individual CFU were then transferred to a multiwell plate containing liquid LB-tet media and...
The adaptation phase for the E. coli culture lasted A culture of E. coli was grown...
The adaptation phase for the E. coli culture lasted
A culture of E. coli was grown on suitable culture media; aliquots of the culture were serially diluted and spread plated aseptically on agar plate media at the times indicated in the table below. Upon overnight incubation at 37°C, colonies appeared and viable counts were colleted and displayed below. Hours 0 Viable Counts (CFU/mL) 2 .00 x 107 2.10 x 107 2.30 x 107 3.25 x 107 6.60 x 107 1.40...
2a. (4 pts) A potent mutagen was added to an E. coli culture growing in rich...
2a. (4 pts) A potent mutagen was added to an E. coli culture growing in rich broth. Prior experiments established that this F cell line, designated WWU113, could not synthesize leucine. You are interested in measuring the rate of induced reverse mutation. You plan to grow a liquid culture of the leu strain and then plate various dilutions of the culture onto selective media. What media should you use to grow up the liquid culture? This will be Media A...
e) Can you identify any wildtype (nonmutant) colonies in the figures? Explain your answer. f) Can...
e) Can you identify any wildtype (nonmutant) colonies in the figures? Explain your answer. f) Can you identify any standard (non-conditional) mutants in the figures? Explain your answer. g) Can you identify any conditional mutants in the figures? Explain your answer. 23°C mutant colony in which cells proliferate at the cooler, permissive temperature but fail to proliferate at the warmer, nonpermissive temperature 23°C colonies replicated onto two identical plates and incubated at two different temperatures mutagenized cells plated out in...
Cloning 2 Below is the restriction map of a 10 kb piece of DNA. Also shown...
Cloning 2
Below is the restriction map of a 10 kb piece of DNA. Also shown
below is a cloning vector which has two unique restriction enzyme
recognition sites, one for EcoRI (E) and one for HindIII (H). The
location of the kanamycin (kan) and ampicillin (amp) resistance
genes is also shown. Kanamycin and ampicillin are antibiotics that
are commonly used to select transformed E. colicells (consult the
Lab Manual for more information). Note that the HindIII site is
located...
2 What is the minimum genotype of a recombinant cell that grew on minimal media supplemented...
2 What is the minimum genotype of a recombinant cell that grew on minimal media supplemented with arginine, methionine and the antibiotic tetracycline but lacking the essential amino acid isoleucine 3 How would you determine if your recombinant cells had also acquired the tryptophan gene from the Hart Complete the following table indicating which of the organisms (F and/or Hfr used in this experiment would be expected to grow on the given media. Unless specified, MMD is just minimal medium...
LAB Genetic Engineering of Bacteria Problem Is it possible to transfer the allele for resistance to...
LAB Genetic Engineering of Bacteria Problem Is it possible to transfer the allele for resistance to the antibiotic ampicillin into a bacterial cell? Objectives After completing this lab, the student will be able to: 1. Demonstrate micropipetting and sterile pipetting techniques for handling and transferring bacteria and plasmid DNA. 2. Maintain sterile conditions for culturing bacterial cells. 3. Inoculate bacteria into flasks, culture tubes, or agar plates. 4. Culture isolated individual colonies from an agar plate to form genetically identical...
2. A dominant allele H reduces the number of body bristles that Drosophila flies have, giving...
2. A dominant allele H reduces the number of body bristles that Drosophila flies have, giving rise to a “hairless” phenotype. In the homozygous condition, H is lethal. An independently assorting dominant allele S has no effect on bristle number except in the presence of H, in which case a single dose of S suppresses the hairless phenotype, thus restoring the "hairy" phenotype. However, S also is lethal in the homozygous (S/S) condition. What ratio of hairy to hairless flies...
Please summarise the following content:(no word limit) EXPERIMENTAL PROCEDURES Animals All animals including pregnant ICR mice,...
Please summarise the following content:(no word
limit)
EXPERIMENTAL PROCEDURES Animals All animals including pregnant ICR mice, neonatal Wistar rats, and NOD-SCID mice (8 weeks old, male) were purchased from CLEA Japan (Tokyo). All exper- imental procedures and protocols were approved by the Animal Care and Use Committee of Keio University and conformed to the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Cells Mouse ESCs were obtained from the Laboratory of Pluripotent Cell Studies, RIKEN...
6. Transposon mutagenesis was performed to identify genes that are involved in the biosynthesis of theA amino acid alanine. A wildtype culture of E. coli was infected with a defective phage carrying Tn10 A and the culture was plated on LB-tet media to obtain tet-resistant colony forming units. (LB is a rich growth media for E. coli and contains all amino acids required for bacterial growth) Individual CFU were then transferred to a multiwell plate containing liquid LB-tet media and...
The adaptation phase for the E. coli culture lasted
A culture of E. coli was grown on suitable culture media; aliquots of the culture were serially diluted and spread plated aseptically on agar plate media at the times indicated in the table below. Upon overnight incubation at 37°C, colonies appeared and viable counts were colleted and displayed below. Hours 0 Viable Counts (CFU/mL) 2 .00 x 107 2.10 x 107 2.30 x 107 3.25 x 107 6.60 x 107 1.40...
2a. (4 pts) A potent mutagen was added to an E. coli culture growing in rich broth. Prior experiments established that this F cell line, designated WWU113, could not synthesize leucine. You are interested in measuring the rate of induced reverse mutation. You plan to grow a liquid culture of the leu strain and then plate various dilutions of the culture onto selective media. What media should you use to grow up the liquid culture? This will be Media A...
e) Can you identify any wildtype (nonmutant) colonies in the figures? Explain your answer. f) Can you identify any standard (non-conditional) mutants in the figures? Explain your answer. g) Can you identify any conditional mutants in the figures? Explain your answer. 23°C mutant colony in which cells proliferate at the cooler, permissive temperature but fail to proliferate at the warmer, nonpermissive temperature 23°C colonies replicated onto two identical plates and incubated at two different temperatures mutagenized cells plated out in...
Cloning 2
Below is the restriction map of a 10 kb piece of DNA. Also shown
below is a cloning vector which has two unique restriction enzyme
recognition sites, one for EcoRI (E) and one for HindIII (H). The
location of the kanamycin (kan) and ampicillin (amp) resistance
genes is also shown. Kanamycin and ampicillin are antibiotics that
are commonly used to select transformed E. colicells (consult the
Lab Manual for more information). Note that the HindIII site is
located...
2 What is the minimum genotype of a recombinant cell that grew on minimal media supplemented with arginine, methionine and the antibiotic tetracycline but lacking the essential amino acid isoleucine 3 How would you determine if your recombinant cells had also acquired the tryptophan gene from the Hart Complete the following table indicating which of the organisms (F and/or Hfr used in this experiment would be expected to grow on the given media. Unless specified, MMD is just minimal medium...
LAB Genetic Engineering of Bacteria Problem Is it possible to transfer the allele for resistance to the antibiotic ampicillin into a bacterial cell? Objectives After completing this lab, the student will be able to: 1. Demonstrate micropipetting and sterile pipetting techniques for handling and transferring bacteria and plasmid DNA. 2. Maintain sterile conditions for culturing bacterial cells. 3. Inoculate bacteria into flasks, culture tubes, or agar plates. 4. Culture isolated individual colonies from an agar plate to form genetically identical...
Please summarise the following content:(no word
limit)
EXPERIMENTAL PROCEDURES Animals All animals including pregnant ICR mice, neonatal Wistar rats, and NOD-SCID mice (8 weeks old, male) were purchased from CLEA Japan (Tokyo). All exper- imental procedures and protocols were approved by the Animal Care and Use Committee of Keio University and conformed to the National Institutes of Health Guide for the Care and Use of Laboratory Animals. Cells Mouse ESCs were obtained from the Laboratory of Pluripotent Cell Studies, RIKEN...
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