how you will choose the concentation of the agarose to see your PCR product .?
Homework Answers
How does agarose gel affect gel electrophoresis ?
Increasing the agarose concentration of a gel reduces the migration
speed and enables separation of smaller DNA molecules. The distance
between DNA bands of a given length is determined by the percent
agarose in the gel.
It all depends on the size of your product. As the greater the % of agarose helps resolve small linear DNA. While lower % gives better resolution and separation of higher molecular-weight bands
Above is the table that show how you can choose the agarose gel
concentration based on the size of DNA separating.
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how
you will choose the concentation of the agarose to see your PCR
product .?
B. To examine whether the PCR was successful, you ran an agarose gel. The expected length of your...
Solution please
B. To examine whether the PCR was successful, you ran an agarose gel. The expected length of your targeted gene was 1,200 base pairs (BP) Lane 10,000 BP | - 4,000 BP 2,000 BP 800 BP : 400 BP A C 1. What were bands A, B, and C? (1 pt)
B. To examine whether the PCR was successful, you ran an agarose gel. The expected length of your targeted gene was 1,200 base pairs (BP) Lane 10,000...
Based on the results from the agarose PCR (240-bp product), how many alanines are likely to...
Based on the results from the agarose PCR (240-bp product), how
many alanines are likely to be inserted in the PHOXB
sequence?
a. 30–40
b. 5–8
c. 15–20
d.
8–12
Case Study 12-2 A 7-month-old child suffered from sleep apnea and difficulty breathing. Respiratory infections and allergies were ruled out. From the family history, the physician suspected congenital congestive hypoventilation syndrome (CCHS). CCHS results, in part, from abnormal function of the PHOX2B gene product caused by a triplet-repeat expansion. The...
When looking at the results of the negative control on the agarose gel, what do you...
When looking at the results of the negative control on the agarose gel, what do you expect to see? Select one: a. The products will be the same size as the blue and brown eye colour samples but the bands visible will be more faint (lighter). b. You will see fewer bands when compared to the blue and brown eye colour samples. c. There should be no PCR product visible. d. You will see a PCR product, but it will...
1. Using your understanding of PCR amplification answer the following questions: a) How do you know...
1. Using your understanding of PCR amplification answer the following questions: a) How do you know if a primer dimer is created during PCR? b) What is nonspecific PCR amplification and how do you know if it exists during PCR? c) What are the possible results you could expect to observe on the agarose gel from your PV92C results? Use a schematic diagram and draw a gel to illustrate where the PV92 primers bind. You should also consider where the...
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...
1. List the ingredients required for PCR. 2. How does DNA move down the agarose gel?...
1. List the ingredients required for PCR. 2. How does DNA move down the agarose gel? What forces aid in this? Meaning, how does gel electrophoresis work? 3. What do the chelex beads do? 4. Why is DNA important?
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in th...
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in this experiment you must add each ingredient separately (unlike the "master-mix" we used in our own experiment). In this experiment, each PCR reaction tube contained the following ingredients added with a micropipetor... 10 HL of 5x Buffer 6 μ1 of 25 mM MgCl2 solution μL of 10 mM dNTP mix 1.5 μL oligonucleotide primer mix 0.5 HL Taq DNA Polymerase stock solution 28...
rate for answering questions compeletly. You plan to use a PCR based assay to determine whether...
rate for answering questions
compeletly.
You plan to use a PCR based assay to determine whether 3 mosquitoes that you have collected are infected with West Nile virus. You have available to you three different virus-specific primers. The primers recognize sequences at the following nucleotide positions on the viral genome: primer 1: 155-176 primer 2: 745-720 primer 3: 926-905 The 5' and 3' ends of the genome are indicated on the long line The first nucleotide at the 5' end...
Explain at least one possible reason why you might have the following results in an agarose...
Explain at least one possible reason why you might have the following results in an agarose gel electrophoresis of your PCR products. In addition to your DNA PCR sample, you also ran a Hi-Lo Marker, a positive PCR control, and a negative PCR control. 16. Scenario C: Your Hi-Lo marker showed up but there were no bands in the lanes for your PCR positive control, DNA PCR sample or negative PCR control.
Can anyone show me step-by-step on how to do the agarose calculations? This week we will...
Can
anyone show me step-by-step on how to do the agarose calculations?
This week we will run the PCR reactions on an agarose gel and analyze the results. Safety notes: Ethidium bromide is a potent mutagen. Wear gloves when handling containers containing ethidium bromide, when handling gels that have been stained in ethidium bromide, and when working with the computer attached to the gel-doc system. Protocol I. Pouring agarose gels I. Using 10x TAE, prepare 50 ml of 3% (w/v)...
Solution please
B. To examine whether the PCR was successful, you ran an agarose gel. The expected length of your targeted gene was 1,200 base pairs (BP) Lane 10,000 BP | - 4,000 BP 2,000 BP 800 BP : 400 BP A C 1. What were bands A, B, and C? (1 pt)
B. To examine whether the PCR was successful, you ran an agarose gel. The expected length of your targeted gene was 1,200 base pairs (BP) Lane 10,000...
Based on the results from the agarose PCR (240-bp product), how
many alanines are likely to be inserted in the PHOXB
sequence?
a. 30–40
b. 5–8
c. 15–20
d.
8–12
Case Study 12-2 A 7-month-old child suffered from sleep apnea and difficulty breathing. Respiratory infections and allergies were ruled out. From the family history, the physician suspected congenital congestive hypoventilation syndrome (CCHS). CCHS results, in part, from abnormal function of the PHOX2B gene product caused by a triplet-repeat expansion. The...
1. List the ingredients required for PCR. 2. How does DNA move down the agarose gel? What forces aid in this? Meaning, how does gel electrophoresis work? 3. What do the chelex beads do? 4. Why is DNA important?
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in this experiment you must add each ingredient separately (unlike the "master-mix" we used in our own experiment). In this experiment, each PCR reaction tube contained the following ingredients added with a micropipetor... 10 HL of 5x Buffer 6 μ1 of 25 mM MgCl2 solution μL of 10 mM dNTP mix 1.5 μL oligonucleotide primer mix 0.5 HL Taq DNA Polymerase stock solution 28...
rate for answering questions
compeletly.
You plan to use a PCR based assay to determine whether 3 mosquitoes that you have collected are infected with West Nile virus. You have available to you three different virus-specific primers. The primers recognize sequences at the following nucleotide positions on the viral genome: primer 1: 155-176 primer 2: 745-720 primer 3: 926-905 The 5' and 3' ends of the genome are indicated on the long line The first nucleotide at the 5' end...
Explain at least one possible reason why you might have the following results in an agarose gel electrophoresis of your PCR products. In addition to your DNA PCR sample, you also ran a Hi-Lo Marker, a positive PCR control, and a negative PCR control. 16. Scenario C: Your Hi-Lo marker showed up but there were no bands in the lanes for your PCR positive control, DNA PCR sample or negative PCR control.
Can
anyone show me step-by-step on how to do the agarose calculations?
This week we will run the PCR reactions on an agarose gel and analyze the results. Safety notes: Ethidium bromide is a potent mutagen. Wear gloves when handling containers containing ethidium bromide, when handling gels that have been stained in ethidium bromide, and when working with the computer attached to the gel-doc system. Protocol I. Pouring agarose gels I. Using 10x TAE, prepare 50 ml of 3% (w/v)...
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