The Qiagen DNeasy DNA extraction kit is used to extract DNA from a liver cell line forsubsequent molecular experiments.(i)(ii)for checking DNA purity using a spectrophotometer.Explain the principle behind this kit in DNA extraction.After the DNA is obtained, the purity of DNA should be determined. Discuss the steps

The Qiagen DNeasy DNA extraction kits are designed for rapid purification of total DNA that is…

Answered: The Qiagen DNeasy DNA extraction kit is…

Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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1) Briefly outline the steps of DNA extraction as carried out in a lab for the purposes of sequencing, with reference to a published protocol from a company that supplies regents and kits for DNA extraction (e.g Qiagen, ThermoFischer, Invitrogen, or similar). Please Include the weblink for the published protocol you used as a reference. (Word limits: 300) 2) Why was there a greater A260 absorbance reading for your DNA sample that was incubated at higher temperatures. (Word Limits: 100)
1) The Qiagen DNeasy DNA extraction kit is used to extract DNA from a liver cell line for subsequent molecular experiments. (i) (ii) Explain the principle behind this kit in DNA extraction. After the DNA is obtained, the purity of DNA should be determined. Discuss the steps for checking DNA purity using a spectrophotometer.
In a protocol for DNA sample preparation for agarose gel electrophoresis, what volume of 4X loading buffer must be added to 21 micro L of DNA to obtain a 1X buffer solution?
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.
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…
Discuss the use of gel electrophoresis for the separation of macromolecules (DNA, RNA and protein) of different sizes and topological forms. (Include the different recombinant DNA techniques that require gel electrophoresis)
Both protein and DNA are run together in an isoelectric focusing (IEF) electrophoresis using the immobilised pH gradient (IPG) strip with pH range of 4-7. After the electrophoresis and staining, only ONE band is observed on the middle of the IPG strip. The band is a protein band. Briefly explain why only the protein band and NOT the DNA band appear on the IPG strip.
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 type of gel(in terms of material) can be more suitable for the electrophoresis of 500-1000bp DNA fragment and why?
Consider the four extraction/purification methods . a. Which extraction method would you use for a touch DNA sample? Explain your reasoning.b. Which extraction method would you use for a sample containing a mixture of primarily non-human and some human DNA? INFO: the 4 extraction methods are QIAamp manual extraction, QIAcube semi-automated extraction, DNA IQ extraction, or FTA punch purification
Q1) a) Draw the absorbance spectra for the following DNA samples. a. Pure DNA without protein contamination DNA with protein contamination b. c. DNA with organic solvent contamination b) An aqueous sample of genomic DNA gave an optical density of 0.6 AU. The sample was diluted 1:50 to achieve an OD below 1.0. The measurement was done in a cuvette with a pathlength of 1 cm. What was the concentration of DNA in the original sample BEFORE dilution? എന്ന് എന്നി per a WANANG KASAMA NA www in de for at de er der er en I
a.) Explain briefly how silica allows us to purify DNA. Be sure to state the relevant biochemical features of silica and DNA as well as any relevant reagents or steps that are important for the purification. Your answer should explain both how we get the DNA to bind in the first place, and how we are able to remove the DNA. You do not (and should not) explain earlier steps in a DNA purification; focus you answer on the silica:DNA interaction. b.) In theory, can a silica spin column also be used to purify RNA? If so, explain how (and why we did not observe RNA when analyzing plasmid DNA via agarose gel electrophoresis). If not, explain why not.

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