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5. If you digested the plasmid above with both EcoRI and HindIII, what would you see?...
1. If a restriction enzyme cuts a circular plasmid twice, how many fragments would you see...
1. If a restriction enzyme cuts a circular plasmid twice, how many fragments would you see on the gel? 2. How would you estimate the total number of base pairs in a plasmid by looking at the DNA fragments of the digested plasmid on a gel? 3. If a linear 1kb DNA fragment has a restriction site that is located 50 bp from one end of the plasmid, what would you expect to see if the digested and undigested DNA...
Now determine the sizes of the fragments generated after digesting the r-plasmid with each of the...
Now determine the sizes of the fragments generated after digesting the r-plasmid with each of the restriction enzymes or combination of restriction enzymes. r-plasmid digested with Pstl r-plasmid digested with EcoRI r-plasmid digested with both Pstl and EcoRI Question 8. (3 pts.) Use this information to determine the relative placement of these restriction sites in the r-plasmid, and then draw restriction site maps of the vector and the r-plasmid on the following circular chromosome templates. The map should show the...
A plasmid is cleaved by restriction endonucleases and analyzed by agarose gel electrophoresis. Assume there is...
A plasmid is cleaved by restriction endonucleases and analyzed
by agarose gel electrophoresis. Assume there is no supercoiling.
Answer the following three questions about the plasmid.
Can you please help below? I also don't understand it so a
explanation for each would be helpful. For the first one I think
its 1000bp but unsure. For the second, I think there is 2 HindIII
sites and for the last one there is 1 EcoRI site? Please
explain.
A plasmid is cleaved...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...
What happens when vector DNA is digested with EcoRI and what does this tell you about...
What happens when vector DNA is digested with EcoRI and what
does this tell you about the number and distribution of EcoRI sites
in the vector?
How big (in base pairs) is (are ) the fragments generated by
digesting the vector with EcoRI?
V R R V & EcoR1 8 , EcoR1, R & & PST1 PST1 Size Markers R & EcoR1 4 Calf Liver Calf & EcoR1 Liver 7 6 5 3 2 1 Wells 16mm Base Pairs 6650...
A. Your lab To see if you understand what you did in our lab, answer the...
A. Your lab To see if you understand what you did in our lab, answer the following based in the procedures for the restriction digest and the gel electrophoresis. 1. Using the PGLO map on p7 of the gel electrophoresis procedure, predict the size of the fragments generated by each enzyme EcoRI Hindill, and Pst! (the sizes you would expect to see on the gel.) (6 pts) Hindill -8 fragments were produced by the restriction enzyme. 2. Answer the calculation...
KpnI (255) lacz alpha gene ECORI (265) Psti (270) PUC origin Smal (3660) рвозобw 3973 bp...
KpnI (255) lacz alpha gene ECORI (265) Psti (270) PUC origin Smal (3660) рвозобw 3973 bp EcoRI (875) Amp(R) Psti (1550) Kan(R) Ncol (1275) 8. You will be using the plasmid shown above as a vector for a DNA cloning experiment. You will create a library of genomic DNA fragments that will then be ligated into this plasmid. The plasmid carries two antibiotic resistance genes, Amp(R) and Kan(R), an origin of replication and a LacZ gene (Lac Z alpha). The...
In your previous prac session you digested your POTC-A plasmid DNA with 3 enzyme mixes (AB...
In your previous prac session you digested your POTC-A plasmid DNA with 3 enzyme mixes (AB and C). One mix contained the restriction endonuclease Kpnl alone, another contained both del and Not and another contained Sphi and Nhe, but you don't know which was which yet. The sites where these enzymes cut POTC-A are indicated on the plasmid map at the top of the page. Calculate the sizes of the DNA fragments that you would expect to see on your...
please help me with these questions Lab 8 Extension Activity: Plasmid Mapping and Restriction Enzymes Mapping...
please help me with these questions
Lab 8 Extension Activity: Plasmid Mapping and Restriction Enzymes Mapping the Plasmid The first step in mapping a plasmid is to determine how many times a restriction site is found on that plasmid. Examine the results for plasmid 55 as an example. The data given in the following table are for the double digest using EcoRI and Pstl. Also, giving are the data for single digests by the individual enzymes. The numbers in the...
8. You are studying a newly identified chromatin-remodeling complex, which you call NICRC. You decide to...
8. You are studying a newly identified chromatin-remodeling complex, which you call NICRC. You decide to run an in vitro experiment to characterize the activity of the purified complex. Your molecular toolbox includes: (1) a 400-base-pair DNA molecule that has a single recognition site for the restriction endonuclease EcoRI, an enzyme that cleaves internal sites on double-stranded DNA (dsDNA); (2) purified EcoRI enzyme; (3) purified DNase I, a DNA endonuclease that will cleave dsDNA at nonspecific sites if they are...
Now determine the sizes of the fragments generated after digesting the r-plasmid with each of the restriction enzymes or combination of restriction enzymes. r-plasmid digested with Pstl r-plasmid digested with EcoRI r-plasmid digested with both Pstl and EcoRI Question 8. (3 pts.) Use this information to determine the relative placement of these restriction sites in the r-plasmid, and then draw restriction site maps of the vector and the r-plasmid on the following circular chromosome templates. The map should show the...
A plasmid is cleaved by restriction endonucleases and analyzed
by agarose gel electrophoresis. Assume there is no supercoiling.
Answer the following three questions about the plasmid.
Can you please help below? I also don't understand it so a
explanation for each would be helpful. For the first one I think
its 1000bp but unsure. For the second, I think there is 2 HindIII
sites and for the last one there is 1 EcoRI site? Please
explain.
A plasmid is cleaved...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...
What happens when vector DNA is digested with EcoRI and what
does this tell you about the number and distribution of EcoRI sites
in the vector?
How big (in base pairs) is (are ) the fragments generated by
digesting the vector with EcoRI?
V R R V & EcoR1 8 , EcoR1, R & & PST1 PST1 Size Markers R & EcoR1 4 Calf Liver Calf & EcoR1 Liver 7 6 5 3 2 1 Wells 16mm Base Pairs 6650...
A. Your lab To see if you understand what you did in our lab, answer the following based in the procedures for the restriction digest and the gel electrophoresis. 1. Using the PGLO map on p7 of the gel electrophoresis procedure, predict the size of the fragments generated by each enzyme EcoRI Hindill, and Pst! (the sizes you would expect to see on the gel.) (6 pts) Hindill -8 fragments were produced by the restriction enzyme. 2. Answer the calculation...
KpnI (255) lacz alpha gene ECORI (265) Psti (270) PUC origin Smal (3660) рвозобw 3973 bp EcoRI (875) Amp(R) Psti (1550) Kan(R) Ncol (1275) 8. You will be using the plasmid shown above as a vector for a DNA cloning experiment. You will create a library of genomic DNA fragments that will then be ligated into this plasmid. The plasmid carries two antibiotic resistance genes, Amp(R) and Kan(R), an origin of replication and a LacZ gene (Lac Z alpha). The...
In your previous prac session you digested your POTC-A plasmid DNA with 3 enzyme mixes (AB and C). One mix contained the restriction endonuclease Kpnl alone, another contained both del and Not and another contained Sphi and Nhe, but you don't know which was which yet. The sites where these enzymes cut POTC-A are indicated on the plasmid map at the top of the page. Calculate the sizes of the DNA fragments that you would expect to see on your...
please help me with these questions
Lab 8 Extension Activity: Plasmid Mapping and Restriction Enzymes Mapping the Plasmid The first step in mapping a plasmid is to determine how many times a restriction site is found on that plasmid. Examine the results for plasmid 55 as an example. The data given in the following table are for the double digest using EcoRI and Pstl. Also, giving are the data for single digests by the individual enzymes. The numbers in the...
8. You are studying a newly identified chromatin-remodeling complex, which you call NICRC. You decide to run an in vitro experiment to characterize the activity of the purified complex. Your molecular toolbox includes: (1) a 400-base-pair DNA molecule that has a single recognition site for the restriction endonuclease EcoRI, an enzyme that cleaves internal sites on double-stranded DNA (dsDNA); (2) purified EcoRI enzyme; (3) purified DNase I, a DNA endonuclease that will cleave dsDNA at nonspecific sites if they are...
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