Homework Answers
- The recombinant DNA, produces two fragments each restriction endonucleases (RENs).
- The total size = 11 kb
- Single PstI digestion = 7 kb + 4 kb
- Double digestion with PstI + EcoRI = 4.3 + 2.7 = 7kb and 3.3 + 0.7 = 4 kb
- Double digestion with PstI + BglI = 6.1 + 0.9 = 7kb and 2.8 + 1.2 = 4 kb
- Thus, EcoRI and BglI sites are within PstI sites.
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Genetics Restriction Mapping
A 7.0kb plasmid is cut at a Pstl site and a 4.0kb piece...
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...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb)....
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
A genetics problem covering chapter 10 concepts. Could really use some help! The Notch gene, involved...
A genetics problem covering chapter 10 concepts. Could
really use some help!
The Notch gene, involved in Drosophila development, is contained within a restriction fragment of Drosophila genomic DNA produced by cleavage with the enzyme SalI. The restriction map of this Drosophila fragment for several enzymes (Sall, PstI, and Xhol) is shown here; numbers indicate the distances between adjacent restriction sites. This fragment is cloned by sticky-end ligation into the single Sall site of a bacterial plasmid vector that is...
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...
B4. Answer ALL parts: The diagram below shows the restriction pattern of a plasmid cut with...
B4. Answer ALL parts: The diagram below shows the restriction pattern of a plasmid cut with the enzymes BamHI, EcoRI and Ndel. The digests are carried out with each enzyme alone and then with different combinations of the three enzymes. Enzyme Fragment sizes kb BamHI 1.4 10.6 EcoRI 4.5 7.5 Ndel 2.5 9.5 BamHI + EcoRI 45 3.4 1.4 BamHI + Ndel 1.4 2.5 5.9 EcoRI + Ndel 0.5 2.0 2.5 7.0 (a) Draw an agarose gel with restriction fragments...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, dige...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below amHI 300 EcoRI 5 kb 3400 1000 EcoRI 2000 Apal Draw the plasmid containing the cloned insert. Indicate clearly where the insert will be located. Include RE...
A restriction map lists the locations of DNA sequences that are cut by a particular restriction...
A restriction map lists the
locations of DNA sequences that are cut by a particular restriction
enzyme for a piece of DNA, such as a chromosome or a plasmid.
Restriction maps are important when generating a construct for
experimental use. Digesting the DNA sequence with the restriction
enzymes will result in fragmented DNA of predictable sizes, based
on the restriction map, that allow a researcher to analyze if his
or her construct was generated correctly when visualized using gel
electrophoresis....
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers...
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers to locations where the restriction endonuclease EcoRI cuts the DNA, and Pst refers to locations where the restriction enzyme Pst cuts the DNA. Potential restriction sites are numbered 1-6. Distances between restriction sites are shown on the bottom scale in base pairs (bp). The thick line represents the part of the molecule that has homology with a probe. Eco Pst Eco Pst Eco Pst...
Chromosomal and plasmid DNA can be cut into manageable pieces by restriction enzymes. Using agarose gel...
Chromosomal and plasmid DNA can be cut into manageable pieces by
restriction enzymes. Using agarose gel electrophoresis, the DNA
fragments can be separated on a gel, based on their lengths. In
order to see the fragments, a stain is typically added to the gel.
The size of each fragment can be determined by comparing each one
to a DNA molecular weight marker of known size.
Below is a map of pBR22 plasmid. The position and base pair
number of the...
A student clones a DNA fragment into the Sall site of pBR322. Sall is a restriction...
A student clones a DNA fragment into the Sall site of pBR322. Sall is a restriction enzyme and pBR322 is the name of a plasmid (diagram below). Predict the media on which she could expect growth of E. coli containing the recombinant plasmid. Explain your answers. (16 pts) 2. EcoRI BamHI Pst Sall Ampicillin Tetracycline resistance (AmpR) resistance (TetR) pBR322 (4,361 bp) Origin of replication (ori) Pvull Media nutrient agar with ampicilin nutrient agar with tetracycline nutrient agar with ampicillin...
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...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
A genetics problem covering chapter 10 concepts. Could
really use some help!
The Notch gene, involved in Drosophila development, is contained within a restriction fragment of Drosophila genomic DNA produced by cleavage with the enzyme SalI. The restriction map of this Drosophila fragment for several enzymes (Sall, PstI, and Xhol) is shown here; numbers indicate the distances between adjacent restriction sites. This fragment is cloned by sticky-end ligation into the single Sall site of a bacterial plasmid vector that is...
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...
B4. Answer ALL parts: The diagram below shows the restriction pattern of a plasmid cut with the enzymes BamHI, EcoRI and Ndel. The digests are carried out with each enzyme alone and then with different combinations of the three enzymes. Enzyme Fragment sizes kb BamHI 1.4 10.6 EcoRI 4.5 7.5 Ndel 2.5 9.5 BamHI + EcoRI 45 3.4 1.4 BamHI + Ndel 1.4 2.5 5.9 EcoRI + Ndel 0.5 2.0 2.5 7.0 (a) Draw an agarose gel with restriction fragments...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below amHI 300 EcoRI 5 kb 3400 1000 EcoRI 2000 Apal Draw the plasmid containing the cloned insert. Indicate clearly where the insert will be located. Include RE...
A restriction map lists the
locations of DNA sequences that are cut by a particular restriction
enzyme for a piece of DNA, such as a chromosome or a plasmid.
Restriction maps are important when generating a construct for
experimental use. Digesting the DNA sequence with the restriction
enzymes will result in fragmented DNA of predictable sizes, based
on the restriction map, that allow a researcher to analyze if his
or her construct was generated correctly when visualized using gel
electrophoresis....
The figure below shows a restriction map of a segment of a DNA molecule. Eco refers to locations where the restriction endonuclease EcoRI cuts the DNA, and Pst refers to locations where the restriction enzyme Pst cuts the DNA. Potential restriction sites are numbered 1-6. Distances between restriction sites are shown on the bottom scale in base pairs (bp). The thick line represents the part of the molecule that has homology with a probe. Eco Pst Eco Pst Eco Pst...
Chromosomal and plasmid DNA can be cut into manageable pieces by
restriction enzymes. Using agarose gel electrophoresis, the DNA
fragments can be separated on a gel, based on their lengths. In
order to see the fragments, a stain is typically added to the gel.
The size of each fragment can be determined by comparing each one
to a DNA molecular weight marker of known size.
Below is a map of pBR22 plasmid. The position and base pair
number of the...
A student clones a DNA fragment into the Sall site of pBR322. Sall is a restriction enzyme and pBR322 is the name of a plasmid (diagram below). Predict the media on which she could expect growth of E. coli containing the recombinant plasmid. Explain your answers. (16 pts) 2. EcoRI BamHI Pst Sall Ampicillin Tetracycline resistance (AmpR) resistance (TetR) pBR322 (4,361 bp) Origin of replication (ori) Pvull Media nutrient agar with ampicilin nutrient agar with tetracycline nutrient agar with ampicillin...
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