Homework Answers
When we add dideoxy adenosine triphosphate i.e., ddATP as it is an abnormal base and it will not alloe the extension of the DNA Further so it will terminate the chain their it self it is one of the method to find out the base pairs and their number
As A pairswith T so when ever we find a T in the sequemce of the plasmid DNA their and their will the dna elongation os terminated and hence the short sequences are as below
31 GCA 51
31 GCAA 51
31 GCAAAC 51
31 GCAAACATTCCT 51
31 GCAAACATTCCTAGC 51
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3. (2 pts.) You are sequencing the end of a genomic DNA fragment that was cut...
Question 13 4 pts A fragment of DNA is cloned into a plasmid with a sequencing...
Question 13 4 pts A fragment of DNA is cloned into a plasmid with a sequencing primer-binding site. After dideoxy sequencing, the gel pattern shown in this diagram is obtained. ddATP reaction ddTTP reaction ddCTP reaction ddGTP reaction What was the sequence of the DNA strand that acted as the template in the sequencing reaction? 5'ACGATCG 3' 5'TGCTAGC 3 5'CGATCGT3 5'GCTAGCA 3'
Suppose you digest the genomic DNA of a particular organism with the restriction enzyme SauA. Then...
Suppose you digest the genomic DNA of a particular organism with
the restriction enzyme SauA. Then you ligate the resulting fragment
into a unique BamHI cloning site of a plasmid. The sequence of the
restriction sites and position of cleavage is shown below
Note: X and Y and their complementary bases Z and Y’
respectively can be any base (A,C, G, or T)
1) As you can see, ligation is possible because the two restriction
enzymes produce compatible sticky ends....
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....
If the target DNA (the 3.266 Kb E. coli genomic Bam HI fragment) has the same restriction sites on each end, there are t...
If the target DNA (the 3.266 Kb E. coli genomic Bam HI fragment)
has the same restriction sites on each end, there are two possible
orientations for the target DNA to insert into the plasmid. The
following restriction enzymes would cut the 3.266 kb Bam HI genomic
fragment containing the RecA gene once or twice or not at all. In
the tables below, list the expected DNA fragment sizes for the two
possible orientations. Round the DNA fragment sizes to...
Easy question Given that a DNA fragment law the end (on right): Which one of the...
Easy question
Given that a DNA fragment law the end (on right): Which one of the following DNA ends is com, Place the following steps in chronological order when creating a tomato genomic DNA library. The available materials are DNA ligase, restriction enzyme, tomato leaves, plasmid DNK to act as vector that has already been digested with the restriction enzyme, competent bacteria, and nutrient agar plates containing ampicillin. You may use a step more than once. Add DNA ligase Transform...
You have cut genomic DNA with AvrII and wish to ligate it into pBluescript. However, there...
You have cut genomic DNA with AvrII and wish to ligate it into pBluescript. However, there is no AvrII site in the multiple cloning region of this plasmid. What other restriction enzyme could you use to cut the plasmid’s multiple cloning region that would produce compatible ends for ligation with your genomic DNA? (Hint: Look up the Compatible Cohesive Ends chart at New England Biolabs website and confer with pBluescript map) (1 point)
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,...
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...
Suppose you want to sequence the following DNA segment: 5’-GATCTCTTGAAATG-primer site-3’ You clone the fragment into...
Suppose you want to sequence the following DNA segment: 5’-GATCTCTTGAAATG-primer site-3’ You clone the fragment into a plasmid, transform it into bacterial cells and isolate DNA from one of the bacterial colonies. You use the Sanger sequencing method to determine the sequence, separating the products from the sequencing reactions by gel electrophoresis. Draw the bands that should appear on the gel from the four sequencing reactions.
You have determined that a bacterial strain you are working with contains a single type of...
You have determined that a bacterial strain you are working with
contains a single type of plasmid. You isolate the plasmid DNA and
digest separate portions of it with each of two different
restriction enzymes, BamHI and HpaI, and also perform a double
digest using both enzymes. You then fractionate the enzyme digests
on an agarose gel and stain the gel with ethidium bromideto
visualize the restriction fragment patterns. Your results are shown
below. Size markers (in nucleotide base pairs)...
Question 13 4 pts A fragment of DNA is cloned into a plasmid with a sequencing primer-binding site. After dideoxy sequencing, the gel pattern shown in this diagram is obtained. ddATP reaction ddTTP reaction ddCTP reaction ddGTP reaction What was the sequence of the DNA strand that acted as the template in the sequencing reaction? 5'ACGATCG 3' 5'TGCTAGC 3 5'CGATCGT3 5'GCTAGCA 3'
Suppose you digest the genomic DNA of a particular organism with
the restriction enzyme SauA. Then you ligate the resulting fragment
into a unique BamHI cloning site of a plasmid. The sequence of the
restriction sites and position of cleavage is shown below
Note: X and Y and their complementary bases Z and Y’
respectively can be any base (A,C, G, or T)
1) As you can see, ligation is possible because the two restriction
enzymes produce compatible sticky ends....
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....
If the target DNA (the 3.266 Kb E. coli genomic Bam HI fragment)
has the same restriction sites on each end, there are two possible
orientations for the target DNA to insert into the plasmid. The
following restriction enzymes would cut the 3.266 kb Bam HI genomic
fragment containing the RecA gene once or twice or not at all. In
the tables below, list the expected DNA fragment sizes for the two
possible orientations. Round the DNA fragment sizes to...
Easy question
Given that a DNA fragment law the end (on right): Which one of the following DNA ends is com, Place the following steps in chronological order when creating a tomato genomic DNA library. The available materials are DNA ligase, restriction enzyme, tomato leaves, plasmid DNK to act as vector that has already been digested with the restriction enzyme, competent bacteria, and nutrient agar plates containing ampicillin. You may use a step more than once. Add DNA ligase Transform...
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,...
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...
You have determined that a bacterial strain you are working with
contains a single type of plasmid. You isolate the plasmid DNA and
digest separate portions of it with each of two different
restriction enzymes, BamHI and HpaI, and also perform a double
digest using both enzymes. You then fractionate the enzyme digests
on an agarose gel and stain the gel with ethidium bromideto
visualize the restriction fragment patterns. Your results are shown
below. Size markers (in nucleotide base pairs)...
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