Homework Answers
22. A positive control tube will be the one which will give an amplified product for sure where as the negative control tube is the one which will not give any product in the end.
A positive control tube contains a template DNA where as the negative control tube does not contain this. Template DNA is chemically synthesized and is complementary to the primers used.
23. The first step of PCR is denaturation which takes place are temperature of around 95 degree Celsius and the duration of this step is 2 minutes per cycle.
The second step of PCR is annealing which takes place at a temperature of around 50 degree Celsius and the duration of this step is 120 seconds per cycle.
The third step of PCR is elongation which takes place at a temperature of around 70 degree Celsius and the duration of this step is also 120 seconds per cycle.
24. In denaturation step, the double stranded DNA helix is denatured at high temperature into single stranded DNA molecules both of which act as template for the synthesis of daughter DNA.
In the second step, annealing, primers bind to 3 prime end of both the strands in a complementary manner.
In the last step, amplification, taq polymerase amplifies the daughter DNA molecule by using primer which have already bind in the annealing step.
25. Number of copies of product formed after n cycles of PCR = 2^n
Here n is 8.
The number of copies of product formed will be 2^8 = 256 copies
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22. What is present in the positive control tube that is not in the negative control...
Description The three steps—the separation of the strands, annealing the primer to the template, and the...
Description The three steps—the separation of the strands, annealing the primer to the template, and the synthesis of new strands—take less than two minutes. Each is carried out in the same vial. At the end of a cycle, each piece of DNA in the vial has been duplicated. The cycle can be repeated 30 or more times, and each newly synthesized DNA piece acts as a new template. After 30 cycles, 1million copies of the initial DNA piece can be...
22-28 Consider the following nucleotide. Sodobno bilololle H ON on | 0-P-0-CH2 O N N CNH,...
22-28 Consider the following nucleotide. Sodobno bilololle H ON on | 0-P-0-CH2 O N N CNH, bring on Ch o osion obi D DO D olabo OH OH MODE a. What is the name of the nucleotide? b. Would this nucleotide be found in both DNA and RNA, only in DNA, or only in RNA? c. What is the name for the type of bond that connects the phosphate and sugar subunits? d. What is the name for the type...
Now. you should be able to answer the following questions: • How the amplification will be...
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En (2 points) You isolated your mitochondrial DNA in Part I. In step 6, you discard...
En (2 points) You isolated your mitochondrial DNA in Part I. In step 6, you discard the supernatant, but keep the pellet. In step 15, you discard the pellet, but keep the supernatant. Explain why the pattern is different between the two steps and the consequence of mixing up these two steps. Procedure Part 1: mt DNA Isolation from your cheek cells. Lysis solution is used to breakdown the cells in this step, you will isolate MEONA from cheek cells....
Can someone help me with the color reactions, positive and negative control, and the 6 questions?...
Can someone help me with the color reactions, positive
and negative control, and the 6 questions?
Sudan IV Test for Lipid 1. Obtain five test tubes and number them 1-5. Your instructor may ask you to test some additional materials. If so, include additional numbered test tubes. 2. To each tube, add the materials listed in Table 3. 3. Add five drops of water to tube 1 and five drops of Sudan IV to each of the remaining tubes. Mix...
12. During the annealing cycle, if the annealing temperature is too low: a. Very low quantity...
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1. Describe the functions of the following reagents in extraction of DNA from corn meal: proteina...
1. Describe the functions of the following reagents in extraction of DNA from corn meal: proteinase K; guanidine HCI; SDS 2. Why is the ratio of the OD at 260 and 280 nm used to estimate DNA purity? 3. In one paragraph, summarize basic principles of PCR technique in your own words. List all the reagents you will need to perform a PCR experiment. Does this method tell you what genetic modifications were made? If yes, describe how you can...
What causes the lack of on in tube # 5 that is acting as the control...
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What would happen if BsteII-HF had an optimal temperature of 30°C but we used the same...
What would happen if BsteII-HF
had an optimal temperature of 30°C but we used the same double
digestion protocol as we used in lab 4? (2pts)
Note: consider all the cloning steps for this
question
Double digestion of the the CAN1.Z ORNA and the PMD2 plasmid Bola I-HF SCHI CANL.Z! CANL.Z -SHI ©- MD2 CANLY → PMD2 CANLLY Bok I-HF You will cut both the plasmid PMD2 and the CAN1.Z ORNA using 2 enzymes (Bstell-HF and Sphl). These are two...
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22-28 Consider the following nucleotide. Sodobno bilololle H ON on | 0-P-0-CH2 O N N CNH, bring on Ch o osion obi D DO D olabo OH OH MODE a. What is the name of the nucleotide? b. Would this nucleotide be found in both DNA and RNA, only in DNA, or only in RNA? c. What is the name for the type of bond that connects the phosphate and sugar subunits? d. What is the name for the type...
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...
En (2 points) You isolated your mitochondrial DNA in Part I. In step 6, you discard the supernatant, but keep the pellet. In step 15, you discard the pellet, but keep the supernatant. Explain why the pattern is different between the two steps and the consequence of mixing up these two steps. Procedure Part 1: mt DNA Isolation from your cheek cells. Lysis solution is used to breakdown the cells in this step, you will isolate MEONA from cheek cells....
Can someone help me with the color reactions, positive
and negative control, and the 6 questions?
Sudan IV Test for Lipid 1. Obtain five test tubes and number them 1-5. Your instructor may ask you to test some additional materials. If so, include additional numbered test tubes. 2. To each tube, add the materials listed in Table 3. 3. Add five drops of water to tube 1 and five drops of Sudan IV to each of the remaining tubes. Mix...
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1. Describe the functions of the following reagents in extraction of DNA from corn meal: proteinase K; guanidine HCI; SDS 2. Why is the ratio of the OD at 260 and 280 nm used to estimate DNA purity? 3. In one paragraph, summarize basic principles of PCR technique in your own words. List all the reagents you will need to perform a PCR experiment. Does this method tell you what genetic modifications were made? If yes, describe how you can...
What causes the lack of on in tube # 5 that is acting as the control tube? 13. What is the reason for using the control tube What is the reason for 5. used in this exercise 14. What is the relationship between the concentration of pepsin and enzyme activity, as observed on Figure 3-7? 15. How does an increase in the amount of the pepsin, in the test tube, enhance the interaction between pepsin and substrate, thus speeding up...
What would happen if BsteII-HF
had an optimal temperature of 30°C but we used the same double
digestion protocol as we used in lab 4? (2pts)
Note: consider all the cloning steps for this
question
Double digestion of the the CAN1.Z ORNA and the PMD2 plasmid Bola I-HF SCHI CANL.Z! CANL.Z -SHI ©- MD2 CANLY → PMD2 CANLLY Bok I-HF You will cut both the plasmid PMD2 and the CAN1.Z ORNA using 2 enzymes (Bstell-HF and Sphl). These are two...
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