Q1)a) Draw the absorbance spectra for the following DNA samples.a. Pure DNA without protein contaminationDNA with protein contaminationb.c. DNA with organic solvent contaminationb) An aqueous sample of genomic DNA gave an optical density of 0.6 AU. The sample wasdiluted 1:50 to achieve an OD below 1.0. The measurement was done in a cuvette with apathlength of 1 cm. What was the concentration of DNA in the original sample BEFOREdilution?എന്ന് എന്നിper aWANANG KASAMA NAwwwinde for at de er der er enI

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Answered: Q1) a) Draw the absorbance spectra for…

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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A 10 μL-aliquot of a resuspended genomic DNA stock solution was obtained and further diluted by adding 990 μL TE buffer. The A260 of the resulting solution was 0.316. Determine its concentration in µg/µL.
Agtergrond / Background: 1. You received four samples of DNA. The first sample was pure DNA dissolved in TE buffer for stability. The other three samples were impure DNA samples. 2. You determined the absorbance at two fixed wavelengths for the pure DNA sample. You obtained the values in Table 1. 3. You determined the absorbance at two fixed wavelengths for the three impure DNA samples. You obtained the values in Table 1. Tabel 1: DNA spektrofotometriese spectrum / Table 1: DNA spectrophotometric Spectrum Wavelength (nm) A260 A280 Sample 1 (OD) 0.186 0.102 Sample 2 (OD) 0.448 0.252 Sample 3 (OD) 0.646 0.357 Sample 4 (OD) 0.196 0.143 4. Determine the 260/280 ratios for each four samples. Record these values in Table 2. Sample 260/280 ratio 1 2 4 Table 2: 260/280 Ratio values for 4 samples of DNA 5. a) Based on these ratios, how pure are the DNA samples you were given? Motivate your answer. 1: 2: 3: 4: b) If a DNA sample is not pure, name one possible contaminant? 6. For sample 1,…
What is the concentration of a DNA solution that absorbs 0.812 and 0.463 at 260 and 280 nm, respectively? Is the DNA solution considered to be good quality? Why or why not?
If the purity of DNA sample is below 1.8 A260/A280, where did the protein contamination come from?
Using a 1 cm cuvette, the absorbance at 260nm of your double-stranded DNA sample is 0.15. What is the concentration of your sample?
Show the calculations for the preparation of 0.01%, 0.001%, 0.0001%, 0.00001% standard DNA solutions via serial dilution from a 0.1% stock solution. Prepare 10.0 mL of 0.10% standard DNA solution using acetate buffer pH 4.6 as solvent. Prepare four different concentrations of the 0.10% standard DNA solution from step 1 via serial dilution to obtain 0.01%, 0.001%, 0.0001%, 0.00001% standard DNA solutions. Obtain seven clean and dry test tubes. Place 1.50 mL solutions in each tube as follows: Table 2.1. Set-up for the diphenylamine assay. Test tube # Content 1 (blank) acetate buffer pH 4.6 2 0.10% standard DNA 3 0.01% standard DNA 4 0.001% standard DNA 5 0.0001% standard DNA 6 0.00001% standard DNA 7 DNA extract from Part I Add 3.50 mL diphenylamine reagent to each tube, swirl each tube to thoroughly mix the contents and heat for 10 mins in a boiling water bath. Cool immediately under tap water. Read absorbances at 595 nm.…
Explain the principles behind DNA isolation using silica membrane spin column from Qiagen. 2 After DNA isolation using Qiagen extraction kit, the final flow through was subjected to spectrophotometric measurements. Explain the situation if the absorbances were found to be: i) The absorbance at 260 nm is higher than absorbance at 280 nm? ii) The absorbance at 230 is higher than absorbance at 260 nm?
Explain how to prepare 2 ml of a solution with a concentration of 1 μg/5ml from a stock solution of a DNA sample with a concentration of 0.1 mg/ml.
Explain how gel filtration chromatography works. What type of gel will you used when the protein size is 2500 Da? Explain.
Isolate B O Isolate A Isolate C O Isolate D The purity and concentration of DNA isolate can be evaluated with the use of UV spectrophotomeleric measurements. measurement of the turbidity of the sample. The organic compounds (containing aromatic Absorbance reading at 320 provides a general rings) used as reagents to extract DNA absorb light strongly at 230. The ideal 260/230 ratio is 2.0-2.2. DNA absorbs light most strongly at 260nm so the absorbance value at this wavelength can be used to estimate the DNA concentration using the equation derived from Beer's Law: Concentration (pg/mL) = (A260 reading -A320 reading) x 50 The absorbance at 280nm is used as an indicator of protein contamination since the aromatic amino acid residues absorb strongly at this wavelength Analyze the data given below and determine the following. Isolate A Isolate B Isolate C Isolate D A320 0.051 0.091 0.065 0.073 A230 1227 1.32 1.95 1.44 A260 4.54 3.92 3.88 4.21 A280 2.01 2.11 2.04 2.32 Question: Which isolate…
Fluorescence confocal microscopy (FCM) - STK38 monoclonal antibody (M01), clone 2G8- 1F3. a. b. 200 μm What is the basic principle of image formation using this microscopy technique? What can be observed and concluded from the image of the specimen?
SHOW COMPLETE SOLUTIONS Q7. A solution of DNA (in TE buffer) gave a A260 = 0.245. The absorbance of the solution at 320 nm is 0.045. What is the concentration of DNA in the solution Q8. From 5 mL gDNA solution, 1 mL aliquot was diluted with sterile DNA gradevwater to a final volume of 5 mL. If the absorbance measured at A260 and A280 are 0.587 and 0.309, respectively (assuming that the solution is not turbid), what is the concentration of DNA sample in the original stock solution?

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Q1) a) Draw the absorbance spectra for the following DNA samples. a. Pure DNA without protein contamination b. DNA with protein contamination c. DNA with organic solvent contamination CORTANA