Add SYBR green to the remaining half of each PCR reaction product and perform a melting curve analysis. slowly increase the temperature of the samples from 60oC to 95oC, recording the fluorescence emitted from each sample with every 5oC increase in temperature. Draw the fluorescence (y axis) versus temperature (x axis) plot that you would expect to see for each product (1 mark each). Specify relevant temperatures for each sample on the x axes of your plots (1 mark each) and provide an explanation to justify your answer for each sample (1 mark each).
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Add SYBR green to the remaining half of each PCR reaction
product and perform a melting...
Can you explain the answers below ? Below is the same gene from page 1. Alternmative...
Can you explain the answers below ?
Below is the same gene from page 1. Alternmative splicing of the primary RNA transcript will include either exon 4 or exon 5. Assume that mutants 1 and 5 are deletion mutants; mutant 2 is a frameshift mutation; and mutant 3, 4, and 6 are substitution mutations. PCR primers are designated with arrows and lowercase letters (a, b, c, d). mutant 4 mutant 61mutant 3 KR Kpnl Kpn ATG TAG TAA exon 1...
2. PCR amplification of the TAS2R38 gene a. The number of copies of the 303 bp sequence grows exp...
2. PCR amplification of the TAS2R38 gene a. The number of copies of the 303 bp sequence grows exponentially (1-2-4-8-etc) after each cycle. The number of cycles we used is on page 97. What is the number of copies of the 303 bp fragment that will theoretically be present at the end of our reaction? b. Denaturation of the 303 bp segment of the TAS2R38 gene is a critical first step in the PCR perties of a DNA segment that...
You have access to a polymerase that has a maximum extension of 3kb. Below is a...
You have access to a polymerase that has a maximum extension of 3kb. Below is a (not to scale) schematic from the paper depicting the location of the transposon within the cct8 gene. For our purposes, the 5' end of Primer 1 is at bp 170 of the WT allele, the 5' end of Primer 2 is at bp 2030 of the WT allele (not including the transposon). The GBT-P9 transposon is 6000 bp long, inserted at bp 300 of...
Please help with all questions. I provided all the information that I have. The sequence below...
Please help with all questions. I provided all the information that I have. The sequence below represents the genomic DNA sequence of the first 440 bp of your gene of interest (exon 1 in blue). You want to amplify this full 440 bp region by PCR, for cloning into a plasmid vector. tgaagtccaactcctaagccagtgccagaagagccaaggacaggtacggctgtcatcacttagacctcaccctgtggagccacaccctagggttggccaatctactcccaggagcagggagggcaggagccagggctgggcataaaagtcagggcagagccatctattgcttacatttgcttctgacacaactgtgttcactagcaacctcaaacagacaccatggtgcatctgactcctgaggagaagtctgccgttactgccctgtggggcaaggtgaacgtggatgaagttggtggtgaggccctgggcaggttgctatcaaggttacaagacaggtttaaggagaccaatagaaactgggcatgtggagacagagaagactcttgggtttctgataggcactgactctctctgcctattggtctattttcccaccc 1.1 Design a 20 nucleotide forward & reverse primer set that will allow you to amplify the sequence above. (note - primers should be at the beginning...
Now. you should be able to answer the following questions: • How the amplification will be...
Now. you should be able to answer the following questions: • How the amplification will be done? - How you will determine your target sequence? How the amplification will be specific for certain segment? What are the requirements to carry PCR? • Suppose you perform a PCR that begins with one double-strand of the following DNA template: +5'-CTACCTGCGGGTTGACTGCTACCTTCCCGGGATGCCCAAAATTCTCGAG-3+ +3'-GATGGACGCCCAACTGACGATGGAAGGGCCCTACGGGTTTTAAGAGCTC-5'+ A. Draw one cycle of PCR reaction below the following diagram. B. Label the template DNA, the primers, and what is...
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....
6. A sequencing chromatograph just arrived! a) Label each peak with an appropriate base (see Olga)...
6. A sequencing chromatograph just arrived! a) Label each peak with an appropriate base (see Olga) b) Highlight mutation(s) site c) Characterize each chromatograph based off mutation(s) you found (give the name(s) of the mutation(s), briefly explain how it would affect the protein. Who is healthy/carrier/affected, why? d) Most importantly: Which mutation did Alexey have in his F9 gene?! e) How did this affect his blood clotting Factor IX? To answer these questions successfully, use the following sequence of the...
alllll them please all To MOST readily demonstrate transformation of bacteria in the laboratory one could...
alllll them please all
To MOST readily demonstrate transformation of bacteria in the laboratory one could extract DNA from: A) his cells and add the DNA to his cells, then grow the cells on plates with histidine B) hist cells and add the DNA to his cells, then grow the cells on plates without histidine C) lac" cells and add the DNA to lact cells, then grow the cells on plates without histidine D) amp cells and add the DNA...
2. A dominant allele H reduces the number of body bristles that Drosophila flies have, giving...
2. A dominant allele H reduces the number of body bristles that Drosophila flies have, giving rise to a “hairless” phenotype. In the homozygous condition, H is lethal. An independently assorting dominant allele S has no effect on bristle number except in the presence of H, in which case a single dose of S suppresses the hairless phenotype, thus restoring the "hairy" phenotype. However, S also is lethal in the homozygous (S/S) condition. What ratio of hairy to hairless flies...
5) Translation was originally sorted out using a variety of inhibitors of protein synthesis Inhibitors tend...
5) Translation was originally sorted out using a variety of inhibitors of protein synthesis Inhibitors tend to interrupt the process at a particular step and a population of translating ribosomes will all stop in the same state. An allied technique is to identify temperature sensitive mutations in various accessory factors. When the population of ribosomes is shifted to the restrictive temperature, ribosomes will stall when they need the function of the mutant protein to continue By analyzing the structure of...
Can you explain the answers below ?
Below is the same gene from page 1. Alternmative splicing of the primary RNA transcript will include either exon 4 or exon 5. Assume that mutants 1 and 5 are deletion mutants; mutant 2 is a frameshift mutation; and mutant 3, 4, and 6 are substitution mutations. PCR primers are designated with arrows and lowercase letters (a, b, c, d). mutant 4 mutant 61mutant 3 KR Kpnl Kpn ATG TAG TAA exon 1...
2. PCR amplification of the TAS2R38 gene a. The number of copies of the 303 bp sequence grows exponentially (1-2-4-8-etc) after each cycle. The number of cycles we used is on page 97. What is the number of copies of the 303 bp fragment that will theoretically be present at the end of our reaction? b. Denaturation of the 303 bp segment of the TAS2R38 gene is a critical first step in the PCR perties of a DNA segment that...
You have access to a polymerase that has a maximum extension of 3kb. Below is a (not to scale) schematic from the paper depicting the location of the transposon within the cct8 gene. For our purposes, the 5' end of Primer 1 is at bp 170 of the WT allele, the 5' end of Primer 2 is at bp 2030 of the WT allele (not including the transposon). The GBT-P9 transposon is 6000 bp long, inserted at bp 300 of...
Now. you should be able to answer the following questions: • How the amplification will be done? - How you will determine your target sequence? How the amplification will be specific for certain segment? What are the requirements to carry PCR? • Suppose you perform a PCR that begins with one double-strand of the following DNA template: +5'-CTACCTGCGGGTTGACTGCTACCTTCCCGGGATGCCCAAAATTCTCGAG-3+ +3'-GATGGACGCCCAACTGACGATGGAAGGGCCCTACGGGTTTTAAGAGCTC-5'+ A. Draw one cycle of PCR reaction below the following diagram. B. Label the template DNA, the primers, and what is...
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....
6. A sequencing chromatograph just arrived! a) Label each peak with an appropriate base (see Olga) b) Highlight mutation(s) site c) Characterize each chromatograph based off mutation(s) you found (give the name(s) of the mutation(s), briefly explain how it would affect the protein. Who is healthy/carrier/affected, why? d) Most importantly: Which mutation did Alexey have in his F9 gene?! e) How did this affect his blood clotting Factor IX? To answer these questions successfully, use the following sequence of the...
alllll them please all
To MOST readily demonstrate transformation of bacteria in the laboratory one could extract DNA from: A) his cells and add the DNA to his cells, then grow the cells on plates with histidine B) hist cells and add the DNA to his cells, then grow the cells on plates without histidine C) lac" cells and add the DNA to lact cells, then grow the cells on plates without histidine D) amp cells and add the DNA...
5) Translation was originally sorted out using a variety of inhibitors of protein synthesis Inhibitors tend to interrupt the process at a particular step and a population of translating ribosomes will all stop in the same state. An allied technique is to identify temperature sensitive mutations in various accessory factors. When the population of ribosomes is shifted to the restrictive temperature, ribosomes will stall when they need the function of the mutant protein to continue By analyzing the structure of...
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