III. Lab review PCR Amplification of a Single DNA Sample Making your Master Mix and PCR Set up for F5 locus Note: For this set, 15 reactions will be conducted in a volume of 15 µL Assume you extract gDNA quantitated by the NanoDrop at a concentration of 1015.3 ng/μl. Calculate the dilution(s) needed to get to a concentration of 25 ng/μL needed for making your Master Mix. Your dNTPs come separately as 100 mM concentrations for ATP, GTP, CTP, and TTP. Calculate the dilution(s) needed to make a dNTP mixture at 2.5 mM for making your Master Mix. Our Forward and Reverse Primers for F5 are resuspended at 100 uM. Calculate the ution(s) needed to make enough primers that can be handled at a pipettable volume for aking your Master Mix, using the desired concentrations on the set up sheet as your guide.

III. Lab review PCR Amplification of a Single DNA Sample Making your Master Mix and PCR Set up for F5 locus Note: For this set, 15 reactions will be conducted in a volume of 15 µL Assume you extract gDNA quantitated by the NanoDrop at a concentration of 1015.3 ng/μl. Calculate the dilution(s) needed to get to a concentration of 25 ng/μL needed for making your Master Mix. Your dNTPs come separately as 100 mM concentrations for ATP, GTP, CTP, and TTP. Calculate the dilution(s) needed to make a dNTP mixture at 2.5 mM for making your Master Mix. Our Forward and Reverse Primers for F5 are resuspended at 100 uM. Calculate the ution(s) needed to make enough primers that can be handled at a pipettable volume for aking your Master Mix, using the desired concentrations on the set up sheet as your guide.

Biology: The Dynamic Science (MindTap Course List)

4th Edition

ISBN:9781305389892

Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Chapter18: Dna Technologies: Making And Using Genetically Altered Organisms, And Other Applications

Section: Chapter Questions

Problem 9TYK

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d. Biuret test O e. NaOH Clear my choice In the method to isolate the DNA from peas, you used the centrifuge right after the addition of salts and alcohol; in which part of the tube will you find the stringy white DNA? of Select one: stion a. Suspended in the solution O b. At this step, no DNA will be visible O c. Top of the tube precipitate O d. Bottom of the tube precipitate Clear my choice
ersonal use only, do not reple 2021-05-12 1ore 7. Restriction endonucleases recognize particular sequences of DNA and then cut the 16@gmail.com DNA into fragments (see table below). RESTRICTION ENDONUCLEASES BamHI DNA SEQUENCE RECOGNIZED* Luse only, do no repro 2021-05 va16@gmail.com GIG ATC C С СТАGJG GỊA A TT C СТТААЈG Pers EcoRI * Į indicates where the DNA backbone is cut The base sequences of DNA from two different individuals, Alice and Ben, are given below. Indicate where BamHI and EcoRI would cut these DNA sequences by drawing lines (for hints refer to FIGURE 2 and the video in the Canvas assignment for this section ). Personal use ofy 2021-05-12 forevermayal6@gmail.com BEN LA ALICE G C G C uCE G C Personal use only. do no G C 2021-05-12 A T Com G C G C forevermaya16@gnT A C G A T ТА C G C G C G G C G C А Т PersA T G C vermaya16@amail.com T A use only, do not reproduce. 2021-05-12 A T А Т А Т ТА ТА ТА C G 7. Does BamHI cut the DNA from Alice and Ben in the same place? C G…
1d) Whether done manually or automated, DNA sequencing gels are always made of polyacrylamide rather than agarose. Why can't agarose be used for a sequencing gel, as it is for other DNA gel electrophoresis?
DNA Extraction by Alkaline Lysis Procedure: 1. Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for 20s to pellet. Remove the supernatant completely with a Pasteur pipet or a plastic pipettor tip. The spins can be performed at FC or at room temperature. Longer spins make it difficult to resuspend cells. 2 Resuspend pellet in 100pul GTE solution and let sit 5 min at room temperature. Be sure cells are completely resuspended. 3. Add 200ul NaOH/SDS solution, mix by tapping tube with finger, and place on ice for 5 min. 4. Add 150ul potassium acetate solution and vortex at maximum speed for 2s to mix. Place on ice for 5-15 min. Be sure mixing is complete. 5. Spin 3 min at 12,000 rpm to pellet cell debris and chromosomal DNA 6. Transfer 0.4 ml supernatant to a fresh tube, mix it with 0.8 ml of 95% ethanol or 0.4 ml isopropanol, and let sit 2 min at room temperature precipitate nucleic acids. 7. Spin at 12,000rpm for 3 min at room temperature to pellet plasmid DNA and…
u+ 2:24 * © . Fundamental of Bio-informa.. Log out DAMINOV AKHMADILLO on 3 In the "Indexing DNA" method of n k-mer table, the maximum number of compares when the k-mers are sorted is ed I out of Select one: g O 1. log2(n) on O 2. nlogn 3. n O 4. None of above 个 TOP
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…
4. Look at the gel image and answer the questions below and be specific. a) Based on your calculations of the DNA concentrations, how much DNA was loaded into each well? Do you see DNA for each of your samples? If not, why do you think that is so? b) Is the DNA in a single sharp band, multiple bands or a smear? What would each of these scenarios be due to, and why would you see them for your samples? c) Do you see multiple bands in your plasmid DNA sample? What are they?
DNA Extraction by Alkaline Lysis Procedure: 1. Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for 20s to pellet. Remove the supernatant completely with a Pasteur pipet or a plastic pipettor tip. The spins can be performed at 4C or at room temperature. Longer spins make it difficult to resuspend cells. 2. Resuspend pellet in 100µl GTE solution and let sit 5 min at room temperature. Be sure cells are completely resuspended. 3. Add 200µl NaOH/SDS solution, mix by tapping tube with finger, and place on ice for 5 min. 4. Add 150µl potassium acetate solution and vortex at maximum speed for 2s to mix. Place on ice for 5-15 min. Be sure mixing is complete. 5. Spin 3 min at 12,000 rpm to pellet cell debris and chromosomal DNA. 6. Transfer 0.4 ml supernatant to a fresh tube, mix it with 0.8 ml of 95% ethanol or 0.4 ml isopropanol, and let sit 2 min at room temperature precipitate nucleic acids. 7. Spin at 12,000rpm for 3 min at room temperature to pellet plasmid DNA and…
1. Definitions and brief statements 1. DNA Data Bank of Japan 2. MeSH Database 3. Please describe the 3 steps and temperature that are cycled during a PCR reaction. ✓ V V 4. What are global alignment and local alignment and their algorithms? ( 5. ProtScale
1. The final amount of each primer required in a PCR reaction is 0.025 nanomol. If the total volume of the PCR reaction is equal to 100 µl and the stock concentration of each primer is equal to 10 µM. Calculate the volume of stock primer that needs to be added in order to ensure a primer amount of 0.025 nanomol. 2. A 250 ml phosphate buffer solution contains 204.12 µg KH2PO4 and 87.09 µg K2HPO4, respectively. Determine the molarities in micromolar (µM) of the abovementioned compounds in the phosphate buffer solution. [Mw KH2PO4 = 136.08 g/mol and Mw K2HPO4 = 174.18 g/mol] 3. You are required to use the pET 28b expression vector in a cloning experiment. If the vector is 5.368 kilobasepairs in length and the mass of the DNA that you have added represent 8.5 x 109 molecules. Determine the mass of vector DNA that you have added to your reaction? [1basepair ~ 660 Da or g/mol, Avogadro’s constant = 6.023 x 1023 molecules/mol]
I. You wish to perform an electrophoretic resolution of your restriction enzyme–digested DNA. The sizes of the expected fragments range from 100 to 500 bp. You discover two agarose gels polymerizing on the bench. One is 0.5% agarose; the other is 2% agarose. Which one might you use to resolve your fragments? I. What does it mean to “denature” a protein and why is this important for SDS PAGE? II. Describe how you denatured the proteins you used for SDS-PAGE. III. Describe the roles of the primary Antibody and the secondary Antibody during an ELISA test. IV. After the addition of the secondary Antibody, how did you determine which wells contain the Antigen bound to primary Antibody bound to secondary Antibody?
Kpnl 4. Plasmid Z has a size of 7 kb, and the map shows Kpnl (K) and Pstl (P) cut sites relative to each other. This plasmid was digested with three different restriction enzymes 2000 bp 3500 bp Kpnl (K), Pstl (P) and Bgll (B) either alone or in combination and the samples run on an agarose gel as shown below. Where does Bgll (B) cut this plasmid ? Does the plasmid have one recognition site or two for Bg|l? Describe the Bgll cut site in this plasmid relative to the Kpnl cut Plasmid Z -7 kb Pstl site. How many bases to the left or right of the Kpnl cut site would you observe the Bgll cut site. Explain briefly. 1500 bp Pstl Ladder Kpni Psti K/P Bgl K/B KPB 7000 bp 7000 bp 5600 bp 5500 bp 4900 bp 3500 bp 2000 bp 1500 bp 1500 bp 1500 bp 1400 bp 600 bp %3D