If your DNA was not cut to completion during the restriction analysis of your transformants, what...
If your DNA was not cut to completion during the restriction analysis of your transformants, what would you expect your gel to look like? Would you still be able to distinguish positive from negative clones?
Homework Answers
DNA separation in agrose gel electrophoresis is occurs based on size and charge of the DNA.
In the given question, restriction digestion of DNA is not complete. In this scenario still, we will able to distinguish positive from negative clones. Incomplete DNA digestion will not result in insert release. But it may lead to nick formation in both positive and negative clones. Irrespective of the status of restriction digestion, the size of positive clones would be larger than negative clones. Differences in size would allow us to distinguish positive and negative clones. This assay called a gel mobility shift assay and frequently used to distinguish positive from negative clones during initial clone screening.
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If your DNA was not cut to completion during the restriction
analysis of your transformants, what...
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....
Are the following statements true or false? Please explain your answers. 1. Restriction nucleases cut DNA...
Are the following statements true or false? Please explain your answers. 1. Restriction nucleases cut DNA at specific sites that are always located between genes. 2. DNA migrates toward the positive electrode during electrophoresis. 3. Clones isolated from cDNA libraries contain promoter sequences. 4. PCR utilizes a heat-stable DNA polymerase because for each amplification step, double-stranded DNA must be heat-denatured.
RESTRICTION DIGEST ANALYSIS QUESTIONS(true or yes = A: false or no = b) 1. Larger DNA...
RESTRICTION DIGEST ANALYSIS QUESTIONS(true or yes = A: false or no = b) 1. Larger DNA fragments appear near the bottom of the gel. 2. Larger DNA fragments move more rapidly through the gel. D ONA that has many restriction sites for a certain endonuclease will be cut into more fragmets than DNA with fewer restriction sites. 4. Cutting DNA with many different endonucleases will result in more DNA fragments. 5. Restriction enzymes all recognize the same base sequence when...
Day 1, You have isolated DNA from 30 individuals and performed restriction enzyme digestion. You have...
Day 1, You have isolated DNA from 30 individuals and performed
restriction enzyme digestion. You have loaded the product and run
the gel electrophoresis following the instruction properly. You
expected to see DNA bands after the procedure is completed, like
figure A, but after the completion of the run your gel looks like
figure B. You did not see any DNA band. You were so upset, but your
lab partner fixed the problem and your gel looked like figure A....
Why do restriction enzymes need to be kept on ice? What order should the DNA, enzyme,...
Why do restriction enzymes need to be kept on ice? What order should the DNA, enzyme, water and buffer be added to the microcentrifuge tube for a restriction digest? If lambda DNA is linear, how many times would the enzyme have to cut the DNA to generate five DNA fragments? Would a shorter DNA fragment move faster or slower through the agarose gel than a longer fragment? Why?
You cut the pBad-mTagBFP2 plasmid with the BamHI restriction enzyme only. There is only one BamHI...
You cut the pBad-mTagBFP2 plasmid with the BamHI restriction enzyme only. There is only one BamHI RE site. How many bands do you expect to see after you run the product on a gel? Would you expect the same result if you cut a linear PCR product that also only has one cut site? Explain your reasoning
1) Imagine that you had two restriction enzymes and a known segment of linear DNA 160...
1) Imagine that you had two restriction enzymes and a known segment of linear DNA 160 kb long: Enzyme A cuts at location(s): 20 kb, 45 kb 70 kb and 110 kb. Enzyme B cuts at location(s): 15 kb and 140 kb. Based on this information, first construct a linear map of this DNA showing the positions of these restriction enzyme cut sites. In the space below.draw what the gel would look like given the following lanes: (8 points total)...
Number of colonies: Plate LB, no DNA LB/ amp, no DNA uncountable LB/ amp, DNA (The two plated lab...
Number of colonies: Plate LB, no DNA LB/ amp, no DNA uncountable LB/ amp, DNA (The two plated labeled Bacteria DNA) 683 (70 ul) 291 (3011) (a) What quantity of DNA (in ng) was used in the transformation? (2 marks) (b) What fraction of the total transformation reaction was plated out? (2 marks) (c) How many transformants (colonies) were there in the total transformation reaction? (2 marks) (d) What was the transformation efficiency, expressed as transformants per ug of DNA...
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...
RECOMBINANT DNA: PLASMID VECTOR engineering is the direct manipulation of an organism's DNA using nology. To...
RECOMBINANT DNA: PLASMID VECTOR engineering is the direct manipulation of an organism's DNA using nology. To begin the recombinant DNA process, scientists must first ide at codes for the production of the protein they want to manufacture. One is to go backwards from the amino acid sequence of the desired protein to ide sequence of the gene. After scientists have identified the gene, they m it. Restriction enzymes or endonucleases from bacterial cells are key in th ia produce 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....
RESTRICTION DIGEST ANALYSIS QUESTIONS(true or yes = A: false or no = b) 1. Larger DNA fragments appear near the bottom of the gel. 2. Larger DNA fragments move more rapidly through the gel. D ONA that has many restriction sites for a certain endonuclease will be cut into more fragmets than DNA with fewer restriction sites. 4. Cutting DNA with many different endonucleases will result in more DNA fragments. 5. Restriction enzymes all recognize the same base sequence when...
Day 1, You have isolated DNA from 30 individuals and performed
restriction enzyme digestion. You have loaded the product and run
the gel electrophoresis following the instruction properly. You
expected to see DNA bands after the procedure is completed, like
figure A, but after the completion of the run your gel looks like
figure B. You did not see any DNA band. You were so upset, but your
lab partner fixed the problem and your gel looked like figure A....
1) Imagine that you had two restriction enzymes and a known segment of linear DNA 160 kb long: Enzyme A cuts at location(s): 20 kb, 45 kb 70 kb and 110 kb. Enzyme B cuts at location(s): 15 kb and 140 kb. Based on this information, first construct a linear map of this DNA showing the positions of these restriction enzyme cut sites. In the space below.draw what the gel would look like given the following lanes: (8 points total)...
Number of colonies: Plate LB, no DNA LB/ amp, no DNA uncountable LB/ amp, DNA (The two plated labeled Bacteria DNA) 683 (70 ul) 291 (3011) (a) What quantity of DNA (in ng) was used in the transformation? (2 marks) (b) What fraction of the total transformation reaction was plated out? (2 marks) (c) How many transformants (colonies) were there in the total transformation reaction? (2 marks) (d) What was the transformation efficiency, expressed as transformants per ug of DNA...
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...
RECOMBINANT DNA: PLASMID VECTOR engineering is the direct manipulation of an organism's DNA using nology. To begin the recombinant DNA process, scientists must first ide at codes for the production of the protein they want to manufacture. One is to go backwards from the amino acid sequence of the desired protein to ide sequence of the gene. After scientists have identified the gene, they m it. Restriction enzymes or endonucleases from bacterial cells are key in th ia produce restriction...
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