Homework Answers
4. The 90 nt ECoRI fragement can be cloned in expression vector and the trucated product thus produced due to frameshift can be a proof for it.
5. To prove a fragment has an IRES, the fragment has to be cloned in vector without a ribosome binding site (RBS) such that any translation from such a fragment can prove it has an internal ribosome entry site (IRES )
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You are given a 90-nt EcoRI fragment of DNA that is theorized to cause a-1 nt...
You have been given a 1.0 kbp human genome fragment that does not contain any repeated...
You have been given a 1.0 kbp human genome fragment that does not contain any repeated sequences. Your plan is to use it as a probe for a Southern Blot containing your friend's genomic DNA. You prepare your gel by digesting with the following restriction enzymes in the following lanes: Lane 1: size marker Lane 2: EcoRI Lane 3: HindIII Lane 4: BamHI Your friend happens to be homozygous for the region you are probing. Does that mean you will...
Luestion 3 1 pts Review: You have the DNA that is radioactively labeled at the S'ends...
Luestion 3 1 pts Review: You have the DNA that is radioactively labeled at the S'ends of DNA as shown below. But you want DNA that is labeled only at one end of the DNA, not both ends. One of the other undergraduate students in the lab suggests that you use a restriction enzyme to cut the DNA, then electrophorese the DNA in an agarose gel, then cut out the region of the gel with radioactive DNA fragment you want,...
A 3.1-kb fragment of DNA containing a complete human gene has been made available to you....
A 3.1-kb fragment of DNA containing a complete human gene has
been made available to you. The DNA segment contains the promoter
sequence and extends beyond the polyadenylation site for the gene.
You decide to do an electromobility shift assay (EMSA) to determine
what molecules bind to the fragment. The results are shown below.
If Lane 1 contains only the 3.1-kb fragment, which answer correctly
describes another lane of the experiment?
a. Lane 2 contains the 3.1kb fragment + TFIIB...
If given a 1.0 kbp human genome fragment that does not contain any repeated sequences. Your...
If given a 1.0 kbp human genome fragment that does not contain any repeated sequences. Your plan is to use it as a probe for a Southern Blot containing your friend's genomic DNA. You prepare your gel by digesting with the following restriction enzymes in the following lanes: Lane 1: size marker Lane 2: EcoRI Lane 3: HindIII Lane 4: BamHI Your friend happens to be homozygous for the region you are probing. Does that mean you will only see...
I have tried but I keep getting them wrong please help!! Thank you! QUESTION 1 Hfr...
I have tried but I keep getting them wrong please help!! Thank you! QUESTION 1 Hfr cells contain the fertility factor as a plasmid, while F+ cells do not. True False QUESTION 2 Hfr cells contain the fertility factor integrated into the bacterial chromosome, while F+ cells contain it as a plasmid. True False QUESTION 3 Both Hfr and F+ cells readily convert F- cells into fertility positive cells. True False 4- Only F+ cells readily convert F- cells into...
1. You are a new undergraduate researcher in the Ruohola-Baker lab. Your job is work with...
1. You are a new undergraduate researcher in the Ruohola-Baker lab. Your job is work with a postdoc named Shiri. Your job is to test the function of a de novo protein named EED binder (EB). In this aim you acquired DNA for this protein. The construct is cloned into pet28b, a bacterial expression vector. Your goal is to express it in human stem cells. A. What method would you use to extract your DNA fragment from the pet28b plasmid?...
1. Fill in the table above with what you observe on your plates. 2. Bacterial transformation...
1. Fill in the table above with what you observe on your
plates.
2. Bacterial transformation occurred on which agar plate (s)?
What evidence do you have that the bacteria were transformed
here?
3. Which plates have glowing growth? Explain what causes
bacteria to glow.
II. Transformation of E. coli with Plasmid DNA (PGLO) 1. Three LB (Luria Broth) agar plates are obtained. The plates contain: • Plate A: LB-Agar/Ampicillin/Arabinose • Plate B: LB-Agar/Ampicillin • Plate C: LB-Agar 2. Three...
1) Imagine that you are interested in studying the enzyme dihydrofolate reductase (DHFR) from Mycobacterium tuberculosis...
1) Imagine that you are interested in studying the enzyme dihydrofolate reductase (DHFR) from Mycobacterium tuberculosis (this enzyme is a potential drug target). The amino acid sequence of the protein is: MTMVGCIWAQATSGVIGRGGDIPWRLPEDQAHFREITMGHTIVMGRRTWDSLPAKVRPLPGRRNWL SRQADFMASGAEWGSLEEALTSPETWVIGGGQVYALALPYATRCEVTEVDIGLPREAGDALAPVLD ETWRGETGEWRFSRSGLRYRLYSYHRS The DNA sequence within the M. tuberculosis genome that codes for the protein is: ATGACGATGGTGGGGCTGATCTGGGCTCAAGCGACATCGGGTGTCATCGGCCGCGGCGGCGACATCCCCT GGCGCTTGCCCGAGGACCAGGCGCATTTCCGGGAGATCACCATGGGGCACACGATCGTGATGGGCCGGCG CACATGGGATTCGCTGCCGGCTAAAGTCCGGCCGCTGCCCGGCCGGCGAAATGTCGTACTGAGCCGCCAA GCTGACTTTATGGCCAGCGGGGCTGAGGTTGTCGGTTCACTCGAGGAGGCGCTGACCAGCCCGGAGACGT GGGTGATCGGAGGCGGACAAGTCTATGCGCTGGCGCTGCCGTACGCGACCAGATGTGAGGTTACCGAGGT CGACATCGGCCTGCCGCGCGAAGCCGGTGACGCGCTGGCCCCCGTGCTGGACGAGACATGGCGGGGCGAG ACGGGGGAGTGGCGCTTCAGCCGGTCCGGGTTGCGGTACCGGTTGTACAGCTACCACCGCTCATGA Assume that you have been given a sample of the bacterial genomic DNA, restriction enzymes Ndel and BamHi, and a plasmid from Novagen called PET-28b....
Your lab instructor has given you a protocol to perform a molecular cloning experiment. In a...
Your lab instructor has given you a protocol to perform a molecular cloning experiment. In a previous experiment, you used polymerase chain reaction (PCR) to amplify a sequence that you believe to regulate expression of a gene you are studying. You will now take this purified PCR product (double stranded DNA) and ligate it into a plasmid that contains a luciferase reporter gene. If your DNA sequence is a promoter sequence, then its presence will allow for expression of the...
pplease explain 24. It was recently (when some of you were high school freshman :-) discovered...
pplease explain
24. It was recently (when some of you were high school freshman :-) discovered that it is possible to obtain information on fetal genotype based on the low number of fetal cells circulating in a pregnant woman's bloodstream. Assume that 1% of all cells in a pregnant woman's bloodstream are from the fetus. You performed high-throughput sequencing of Jane Doe's blood, and for the beta globin gene obtained the following results: 5'-ACTCCTGAGGAGAAG-3' 5'-ACTCCTGTGGAGAAG-3' (wild type allele) (Hbbs allele)...
Luestion 3 1 pts Review: You have the DNA that is radioactively labeled at the S'ends of DNA as shown below. But you want DNA that is labeled only at one end of the DNA, not both ends. One of the other undergraduate students in the lab suggests that you use a restriction enzyme to cut the DNA, then electrophorese the DNA in an agarose gel, then cut out the region of the gel with radioactive DNA fragment you want,...
A 3.1-kb fragment of DNA containing a complete human gene has
been made available to you. The DNA segment contains the promoter
sequence and extends beyond the polyadenylation site for the gene.
You decide to do an electromobility shift assay (EMSA) to determine
what molecules bind to the fragment. The results are shown below.
If Lane 1 contains only the 3.1-kb fragment, which answer correctly
describes another lane of the experiment?
a. Lane 2 contains the 3.1kb fragment + TFIIB...
1. You are a new undergraduate researcher in the Ruohola-Baker lab. Your job is work with a postdoc named Shiri. Your job is to test the function of a de novo protein named EED binder (EB). In this aim you acquired DNA for this protein. The construct is cloned into pet28b, a bacterial expression vector. Your goal is to express it in human stem cells. A. What method would you use to extract your DNA fragment from the pet28b plasmid?...
1. Fill in the table above with what you observe on your
plates.
2. Bacterial transformation occurred on which agar plate (s)?
What evidence do you have that the bacteria were transformed
here?
3. Which plates have glowing growth? Explain what causes
bacteria to glow.
II. Transformation of E. coli with Plasmid DNA (PGLO) 1. Three LB (Luria Broth) agar plates are obtained. The plates contain: • Plate A: LB-Agar/Ampicillin/Arabinose • Plate B: LB-Agar/Ampicillin • Plate C: LB-Agar 2. Three...
1) Imagine that you are interested in studying the enzyme dihydrofolate reductase (DHFR) from Mycobacterium tuberculosis (this enzyme is a potential drug target). The amino acid sequence of the protein is: MTMVGCIWAQATSGVIGRGGDIPWRLPEDQAHFREITMGHTIVMGRRTWDSLPAKVRPLPGRRNWL SRQADFMASGAEWGSLEEALTSPETWVIGGGQVYALALPYATRCEVTEVDIGLPREAGDALAPVLD ETWRGETGEWRFSRSGLRYRLYSYHRS The DNA sequence within the M. tuberculosis genome that codes for the protein is: ATGACGATGGTGGGGCTGATCTGGGCTCAAGCGACATCGGGTGTCATCGGCCGCGGCGGCGACATCCCCT GGCGCTTGCCCGAGGACCAGGCGCATTTCCGGGAGATCACCATGGGGCACACGATCGTGATGGGCCGGCG CACATGGGATTCGCTGCCGGCTAAAGTCCGGCCGCTGCCCGGCCGGCGAAATGTCGTACTGAGCCGCCAA GCTGACTTTATGGCCAGCGGGGCTGAGGTTGTCGGTTCACTCGAGGAGGCGCTGACCAGCCCGGAGACGT GGGTGATCGGAGGCGGACAAGTCTATGCGCTGGCGCTGCCGTACGCGACCAGATGTGAGGTTACCGAGGT CGACATCGGCCTGCCGCGCGAAGCCGGTGACGCGCTGGCCCCCGTGCTGGACGAGACATGGCGGGGCGAG ACGGGGGAGTGGCGCTTCAGCCGGTCCGGGTTGCGGTACCGGTTGTACAGCTACCACCGCTCATGA Assume that you have been given a sample of the bacterial genomic DNA, restriction enzymes Ndel and BamHi, and a plasmid from Novagen called PET-28b....
pplease explain
24. It was recently (when some of you were high school freshman :-) discovered that it is possible to obtain information on fetal genotype based on the low number of fetal cells circulating in a pregnant woman's bloodstream. Assume that 1% of all cells in a pregnant woman's bloodstream are from the fetus. You performed high-throughput sequencing of Jane Doe's blood, and for the beta globin gene obtained the following results: 5'-ACTCCTGAGGAGAAG-3' 5'-ACTCCTGTGGAGAAG-3' (wild type allele) (Hbbs allele)...
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