Homework Answers
First parameter is the GC content of the strand. The more the GC content the more is the the temperature required for denaturation of the two strands.
Second we must make sure that there are not secondary structures in the the gene or else faulty or no amplification will occur
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(4 pts) You are designing new PCR primers for MLST and need to identify a gene...
4. (1.5 pts)You are practicing designing primers that you can use in PCR reactions. You want...
4. (1.5 pts)You are practicing designing primers that you can use in PCR reactions. You want your primers to allow you to amplify the sequence found below. 5°-АсттсслтАTстсТАЛААТАССАТCGATСтстсссссстАGстAСCТААССАGAGACCСТАССG-3. 3-тслАсстатАсAGATTTTATсстасстаGлCAссссССATCCATCGAтTСстстстасGAТСCС-5. Markers part (a) part (b) part ck Right primer should anneal to this region Left primer should 87 nts anneal to this region 80 nts 70 nts 60 nts 50 nts 40 nts 27 nts Draw into the following gel lanes what size(s) of PCR products you would get if you...
1. Create the primers to amplify the gene of interest. 1. Below is almost the full...
1. Create the primers to amplify the gene of interest. 1. Below is almost the full sequence of the coding strand of the F9 gene (exon 2-exon4) that you found in the online database (NCBI, Genomic sequence F9 gene). Some of the sequence is missing, but you know how many nucleotides are skipped. Design forward and reverse primers 18nt each for PCR that will isolate EXACTLY the sequence that is in bold typeface. Exon 2 = 164 nt Intron 2...
You’re trying to PCR amplify a gene, but your gel shows that there are two bands...
You’re trying to PCR amplify a gene, but your gel shows that there are two bands instead of only one like you expected. You know for certain that there no homologous genes in the genome from which you’re amplifying, so you assume that the issue must be coming from your primers or PCR conditions. Briefly describe 2 different strategies for solving the problem and how they work.
You decide to use PCR to determine if your genotype for the PTC tasting gene TAS2R38....
You decide to use PCR to determine if your genotype for the PTC tasting gene TAS2R38. You then decide to also determine genotype for another gene called PER3. What PCR "ingredient" would be different in these two tests? Select one: a. Template gDNA b. dNTPS (nucleotides) c. Taq DNA polymerase d. Primers
1. What are the degenerate primers in the PCR amplification protocol? What do they do? 2....
1. What are the degenerate primers in the PCR amplification protocol? What do they do? 2. Suppose you begin a PCR reaction with 1 piece of double stranded DNA. After 30 cycles of replication, how many pieces of double stranded DNA do you now have? 3. What would be the consequence of having too much DNA in the sample? Would it interfere with the PCR reaction? Why? 4. What happens during the annealing step of the PCR reaction? During the...
• 4. How many copies of the primers do you add to the PCR? You add...
• 4. How many copies of the primers do you add to the PCR? You add 3 pl. of a 5 M solution of each primer, If you had 100 copies of the template DNA would there be enough primers to support 30 cycles of PCR? Remember the amount of template doubles (in theory) with each cycle. After 1 cycle there will be 2xthe starting amount, after 2 cycles 4 x the starting amount.
You are interested in cloning a gene from the B. sanfranciscus genome, so you design PCR...
You are interested in cloning a gene from the B. sanfranciscus genome, so you design PCR primers that should amplify a 1 kilobase pair (kbp) PCR product that contains the gene of interest. After amplification, you will see if the PCR was successful by loading the entire reaction onto an agarose gel and performing electrophoresis to see if a product of the expected size was generated. To visualize the DNA, you will stain the gel with a fluorescent dye called ethidium bromide, which...
Primers are short pieces of DNA that are used in PCR (Polymerase Chain Reaction), the process...
Primers are short pieces of DNA that are used in PCR (Polymerase Chain Reaction), the process which is used in molecular labs to make copies of short stretches of a gene or genome. You have designed primes that have a Molecular Weight (MW) of 6000. It arrives as a dry sample in the quantity of 300 µg. How much water do you need to add to make a primer stock solution of 100 mM primers? HINTS: Remember that mM is...
Instructions: Design primers for genotyping a 200bp deletion in gene Y6B3B.10 (WIT 7470 bp). Copy and...
Instructions: Design primers for genotyping a 200bp deletion in gene Y6B3B.10 (WIT 7470 bp). Copy and Paste in the space below deletion in gene Lng-1gk327) Q1. (2 pts) Annotate (highlight) your primers. Provide your sequences below in the provided space as needed for ordering. orward Q2. (I pt) In a single sentence explain what the functional group must be on your primer and what is its purpose? the 3' end of Q3.(1 pt) List 4 key properties that your primers...
Instructions: Design primers for genotyping a 200bp deletion in gene Y6B3B.10 (WIT 7470 bp). Copy and...
Instructions: Design primers for genotyping a 200bp deletion in gene Y6B3B.10 (WIT 7470 bp). Copy and Paste in the space below deletion in gene Lng-1gk327) Q1. (2 pts) Annotate (highlight) your primers. Provide your sequences below in the provided space as needed for ordering. orward Q2. (I pt) In a single sentence explain what the functional group must be on your primer and what is its purpose? the 3' end of Q3.(1 pt) List 4 key properties that your primers...
4. (1.5 pts)You are practicing designing primers that you can use in PCR reactions. You want your primers to allow you to amplify the sequence found below. 5°-АсттсслтАTстсТАЛААТАССАТCGATСтстсссссстАGстAСCТААССАGAGACCСТАССG-3. 3-тслАсстатАсAGATTTTATсстасстаGлCAссссССATCCATCGAтTСстстстасGAТСCС-5. Markers part (a) part (b) part ck Right primer should anneal to this region Left primer should 87 nts anneal to this region 80 nts 70 nts 60 nts 50 nts 40 nts 27 nts Draw into the following gel lanes what size(s) of PCR products you would get if you...
1. Create the primers to amplify the gene of interest. 1. Below is almost the full sequence of the coding strand of the F9 gene (exon 2-exon4) that you found in the online database (NCBI, Genomic sequence F9 gene). Some of the sequence is missing, but you know how many nucleotides are skipped. Design forward and reverse primers 18nt each for PCR that will isolate EXACTLY the sequence that is in bold typeface. Exon 2 = 164 nt Intron 2...
1. What are the degenerate primers in the PCR amplification protocol? What do they do? 2. Suppose you begin a PCR reaction with 1 piece of double stranded DNA. After 30 cycles of replication, how many pieces of double stranded DNA do you now have? 3. What would be the consequence of having too much DNA in the sample? Would it interfere with the PCR reaction? Why? 4. What happens during the annealing step of the PCR reaction? During the...
• 4. How many copies of the primers do you add to the PCR? You add 3 pl. of a 5 M solution of each primer, If you had 100 copies of the template DNA would there be enough primers to support 30 cycles of PCR? Remember the amount of template doubles (in theory) with each cycle. After 1 cycle there will be 2xthe starting amount, after 2 cycles 4 x the starting amount.
Instructions: Design primers for genotyping a 200bp deletion in gene Y6B3B.10 (WIT 7470 bp). Copy and Paste in the space below deletion in gene Lng-1gk327) Q1. (2 pts) Annotate (highlight) your primers. Provide your sequences below in the provided space as needed for ordering. orward Q2. (I pt) In a single sentence explain what the functional group must be on your primer and what is its purpose? the 3' end of Q3.(1 pt) List 4 key properties that your primers...
Instructions: Design primers for genotyping a 200bp deletion in gene Y6B3B.10 (WIT 7470 bp). Copy and Paste in the space below deletion in gene Lng-1gk327) Q1. (2 pts) Annotate (highlight) your primers. Provide your sequences below in the provided space as needed for ordering. orward Q2. (I pt) In a single sentence explain what the functional group must be on your primer and what is its purpose? the 3' end of Q3.(1 pt) List 4 key properties that your primers...
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