To be able to properly analyze a sample of DNA, you need to have at least 1 µg of the DNA of interest. Assumptions: PCR is run with 30 cycles; Molecular weight per nucleotide in DNA is about 660 g/bp (330 g/bp for single stranded DNA or RNA); bp = base pair. a. Given that you can use the Polymerase Chain Reaction (PCR) to amplify DNA from the original sample and the size of the fragment to be amplified is 10,000 nucleotides long, estimate how many DNA molecules you need to obtain the minimum amount required to analyze the sample. b. Suggest a way to increase your sensitivity, i.e. amplify the DNA starting from even fewer molecules of template DNA.

To be able to properly analyze a sample of DNA, you need to have at least 1 µg of the DNA of interest. Assumptions: PCR is run with 30 cycles; Molecular weight per nucleotide in DNA is about 660 g/bp (330 g/bp for single stranded DNA or RNA); bp = base pair. a. Given that you can use the Polymerase Chain Reaction (PCR) to amplify DNA from the original sample and the size of the fragment to be amplified is 10,000 nucleotides long, estimate how many DNA molecules you need to obtain the minimum amount required to analyze the sample. b. Suggest a way to increase your sensitivity, i.e. amplify the DNA starting from even fewer molecules of template DNA.

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter15: Genomes And Genomics

Section: Chapter Questions

Problem 6QP: Which of the following best describes the process of DNA sequencing? a. DNA is separated on a gel,...

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a) (iii) (iv) Polymerase chain reaction (PCR) is a technique that enables multiplication of specific DNA sample at minute amount to millions or billions of copies at a short time span. (i) Figure 1 indicates the chemicals added into the PCR reaction tube prior to the addition of thermostable DNA polymerase. Do you agree with the list? Justify your answer. DNA template Buffer (containing Tris-HCI, KCI, Mg2+) ddNTP Forward primer Reverse primer Figure 1 If TA cloning is planned to be carried out after amplification of the gene, which thermostable DNA polymerase will you select and the reason for your selection? Why is the optimal annealing temperature vital in this technique? Explain how this temperature (too high or too low) will affect the efficiency of this reaction. If the primers you purchased possessed the following information: Number of Guanine: 4 Number of Adenine: 3 Number of Thymine: 4 Number of Cytosine: 5 Calculate the melting temperature of this primer and estimate the…
PCR is a molecular biology technique where template DNA is amplified using a primer and oligonucleotides. The reaction is catalyzed by a thermostable DNA polymerase and in a particular reaction, the template strands are denatured at 95˚C. For strand hybridization, the melting temperature is 55˚C. What do you predict about the average duration of H bonds at the high temperature in comparison to the low temperature?
Which of the following best describes the process of DNA seqencing. a. DNA is seperated on a gel and the different bands are labled with flouroscent nucleotides and scanned with a laser. b. A laser is used to flurorescently label the nucleotides present with in the DNA , the DNA is run on a gel and then the DNA is droken into fragments c. Nucleotides are scanned with a laser and incrprorated into the DNA that has been seperated on a gel and then DNA is amplified with PCR. d. fragments of DNA are produced in a reaction that lables them with any of four different fluroscent dyes and the fragmented then are run on a gel and scanned with laser e. DNA is broken down into its constituents nucleotides and the nucleotides are then run on a gel and purified with a laser
Polymerase chain reaction (PCR) is a technique that enables multiplication of specific DNA sample at minute amount to millions or billions of copies at a short time span. (i) (ii) Figure 1 indicates the chemicals added into the PCR reaction tube prior to the addition of thermostable DNA polymerase. Do you agree with the list? Justify your answer. DNA template Buffer (containing Tris-HCI, KCI, Mg2+) ddNTP Forward primer Reverse primer Figure 1 If TA cloning is planned to be carried out after amplification of the gene, which thermostable DNA polymerase will you select and the reason for your selection?
Why is the company Qiagen has more refined DNA extraction steps than a normal Strawberry DNA extraction practical? Summary of Qiagen DNA extraction steps Add ATL buffer and grind with sample. Add 20 microliters of enzyme Proteinase K to degrade protein into a 1.5-2ml microcentrifuge tube. Add 200 microlitres AL lysis buffer, and mix by vortexing for 5–10 seconds, which breaks cell membrane allowing DNA to be released. Incubate the sample at 56 degrees for 10 minutes. Mix the cell lysate with 200 microlitres ethanol by pipetting it at the side of the microcentrifuge wall so DNA precipitates. The DNA forms a white layer and the remaining liquid is discarded. Pipet the mixture into DNeasy Mini spin column placed in a 2 ml collection tube. Centrifuge for a minute at 8000 rpm. Place the mini spin column into a 2 ml collection tube, add 500 µl Buffer AW1, and centrifuge for 1 min at 8000 rpm. Then add it to a new 2 ml collection tube (provided), add 500 µl Buffer AW1, and centrifuge for 1…
Which of the following chemicals or procedures is not involved in a typical Polymerase Chain Reaction (PCR). Select one: O a. ddNTPs b. O C. O d. Oe. O f. Repeated cycling (typically 32 cycles) for exponential target amplification DNA strand replication/extension Annealing of oligonucleotide primer or primers Thermostable (Taq) DNA polymerase Buffer with required cofactor: Mg 2+ Og. DNA template Oh. High temperature denaturation O i. dNTPs
Overall, a molecule of DNA does not have a positive or negtive charge since the bases are hydrophobic. Select one: True False You loaded a gel with DNA size standards and then 3 different PCR samples. After running the gel until the Bromophenol blue was near the end of the gel, you put the gel under UV light BUT saw no bands at all in any of the 4 lanes on the gel. What could be the problem? a. The DNA fragments must have run off the end of the gel. b. Your PCR reactions did not work. c. All of the answers presented here could be the problem d. The leads (wires) from the power source were placed backwards on the gel unit. e. You forgot to add ethidium bromide to the gel.
All of the following are examples of materials that are bound by your purifying medium during the DNA extraction process, except: O endoplasmic reticulum extraneous DNA O Calcium (Ca2+) O Iron (Fe2+) O Magnesium (Mg2+) O lipid membranes
You digest 4 uL of plasmid DNA that is 50 ng/uL concetration in a total volume of 20 uL. You run 10 uL of the digest on teh gel. You then do a DNA purification protocol with a Zippy prep on the remaining digested DNA. You elute the DNA in a 25 uL. A 2 uL ssample has the concentration of 2 ng/uL. What is the DNA yield? YIELD = How much dna came out of the column/how much DNA was loaded onto the columnRemember to subtract amount of DNA run on gel from total digested DNA to get hw much DNA was loaded onto the columnAmount that came out of the column equals the volume of the eluted DNA times the concentration of the purified DNA
You are performing PCR for the first time using some new primers that are 25 nucleotides in length with an estimated melting temperature of 65 0C to amplify a 200 nucleotide gene. After PCR, you run an agarose gel on the samples and observe a faint band at 25 nucleotides. What is the best possible explanation for the results? The primers hybridized to multiple sites on the DNA template. Too much template DNA was added to the PCR mixture. The primers dimerized preventing DNA transcription from occurring. A nuclease was in the solution causing degradation of the DNA.
You digest 4 uL of plasmid DNA that is 50 ng/uL concetration in a total volume of 20 uL. You run 10 uL of the digest on teh gel. You then do a DNA purification protocol with a Zippy prep on the remaining digested DNA. You elute the DNA in a 25 uL. A 2 uL ssample has the concentration of 2 ng/uL. What is the DNA yield?
The polymerase chain reaction (PCR) is used by scientists to amplify DNA, particularly when the quantity of DNA is very small, mixed, or contaminated with other organisms. Explain (with words) the how PCR works using a diagram to help illustrate this (show a minimum of three cycles to illustrate your point).