Please help solve these. I have tried for over an hour and I am not getting anywhere on any of these II.Making your Master Mix and PCR Set up for TPCN2:Note: the reaction volumes will be 20 uL and n10 reactions!Assume you extract gDNA quantitated by the NanoDrop at a concentration of 1065.7 ng/μL.Calculate the dilution(s) needed to get to a concentration of 25 ng/μL needed for making yourMaster Mix. Show pipettable volumes! Do two-step dilution.... Show work.Your dNTPs come separately as 100 mM concentrations for ATP, GTP, CTP, and TTP. Showwhat is mixed at what volume. Calculate the dilution (s) needed to make a dNTP mixture at2.5 mM for making your Master Mix. Make sure to prepare enough for all your reactions.Your Forward and Reverse Primers for TPCN2 are re-suspended for you at 100 uM.Calculate the dilution(s) needed to make enough primers that can be handled at a pipettablevolume for making your Master Mix, using the desired concentrations on the set up sheet.

Genomic deoxyribonucleic acid (abbreviated as gDNA) represents chromosomal DNA. This DNA is in…

Note: the reaction volumes will be 20 μL and n = 10 reactions! Assume you extract gDNA quantitated by the NanoDrop at a concentration of 1065.7 ng/μL. Calculate the dilution(s) needed to get to a concentration of 25 ng/μL needed for making your Master Mix. Show pipettable volumes! Do two-step dilution.... Show work.

Note: the reaction volumes will be 20 μL and n = 10 reactions! Assume you extract gDNA quantitated by the NanoDrop at a concentration of 1065.7 ng/μL. Calculate the dilution(s) needed to get to a concentration of 25 ng/μL needed for making your Master Mix. Show pipettable volumes! Do two-step dilution.... Show work.

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

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1) Prepare the following enzymatic reaction, present it in tabulated form. In a final volume of 30 ul, where buffer 4 (10 ml). How much volume of each reagent would be used and how much of water? Is there any problem? 2) The DNA pol 1 enzyme comes at a concentration of 50,000 U/ml. You have to prepare a 50 ug PCR reaction where you must use 0.05 U/ml reaction. You add 10 ul of PCR buffer, 2 ng of tempered DNA that is at a concentration of 0.5 ng/ul, primers (which are at 200 mM) so that each one remains at a concentration of 200 uM, Mg+2 that is 5 mM (10 X), enzyme and water. Present the table of all the reagents included in the reaction, the volumes of each one in ul. Present where the initial and final concentration of each reagent applies. Assume you have micropipettes for all values.
Note: Answer the number 3 question only. Explain. 1. What is the Difference between DNA Purification and RNA Purification? 2. why contaminants must be avoided in DNA samples? 3. Give 5 examples of sources of error in using a spectrophotometer.
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…
Note:Answer the number 2 question only... 1. What is the Difference between DNA Purification and RNA Purification? 2. why contaminants must be avoided in DNA samples? 3. Give 5 examples of sources of error in using a spectrophotometer.
3 Step3: After you have prepared the 3 diluted plasmid samples and the master mix, you can now use them to prepare the individual PCR reactions. For example, to prepare 25 μl PCR reactions for sample A, you need to combine, J ul of "diluted plasmid sample A", and M ul of "master mix". Repeat this step for all 3 reactions, then you can start the PCR reaction in the thermocycler. Solve for J and M.
In bacterial transformation ... 1, Why do you have to incubate the competent bacterial cells on ice after the addition of plasmid DNA? 2. Why do you have to add SOC (Super Optimal Broth with Catabolite repression) medium to bacterial cells? 3. Why do you have to incubate the bacterial cells in SOC medium at 37oC for 45 minutes?
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
1. Compute the volumes needed to complete this table. Stock Final Volume for Volume for 5x reaction Reaction Component Concentration Concentration 1x reaction sddH,0 up to 25 ul up to 25 ul Buffer (ViBuffer A) 10X 1X MgCl, 50 mM 3 mM DNTP 2 mM 100 uM forward primer 10 uM 0.2 uM reverse primer 10 uM 0.2 uM Taq polymerase 1 U/ ul 1 U/ 25 ul DNA template 1-2 ng 1 ul *5X reactions = 2 samples, 1 NTC, 1 positive control, 1 pipetting allowance
Equilibrium Density Gradient Centrifugation of DNA Samples Review the following terms using the internet before working on the problem: radioactive labeling, DNA, cesium chloride, equilibrium centrifugation, DNA denaturation using heat EΧPERIΜΕT Radioactively labeled bacterial DNA was analyzed by cesium chloride (CSCI) equilibrium density gradient centrifugation. The radioactivity of fractions collected from the gradients, and the densities of the two peak fractions, were determined. Graph A: Native DNA sample; Graph B: DNA sample heated to 100°C and rapidly cooled on ice before centrifugation.
Could you please explain these: 1. Why analyze the unknowns in a (short) serial dilution rather than at a single concentration? 2. Which technique can detect lower concentrations of DNA? Compare the detection limit of the fluorescence approach and the absorbance approach
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?
Can you please help with 1b please. picture with 1 graph is for question 1a) picture with 4 graphs is for question 1b) 1a) E. coli DNA and binturong DNA are both 50% G-C. If you randomly shear E. coli DNA into 1000 bp fragments and put it through density gradient equilibrium centrifugation, you will find that all the DNA bands at the same place in the gradient, and if you graph the distribution of DNA fragments in the gradient you will get a single peak (see below). If you perform the same experiment with binturong DNA, you will find that a small fraction of the DNA fragments band separately in the gradient (at a different density) and give rise to a small "satellite" peak on a graph of the distribution of DNA fragments in the gradient (see below). Why do these two DNA samples give different results, when they're both 50% G-C? 1b) If you denatured the random 1000 bp fragments of binturong DNA that you produced in question 1a by heating them to 95ºC, and then cooled them down to 60ºC…