r DNA isolation using Qiagen extraction kit, the ophotometric measurements. Explain the situation i) The absorbance at 260 nm is higher than absort
Q: Ethidium bromide used to be a common stain for detecting DNA/ RNA on agarose gel. (i) Describe how…
A: Ethidium Bromide is a DNA stain as well as a mutagen. This dye can be visualized under UV light. It…
Q: Give advantages and disadvantages of using commercially available kits for plasmid extraction over…
A: As molecular biology techniques get more sensitive, a DNA extraction kit that yields high-quality…
Q: DNa Mapping using Restriction enzymes lab: We will be aliquoting and delivering 5 μl of enzyme to…
A: A restriction enzyme, also known as restriction endonuclease is an enzyme that cleaves DNA into…
Q: Hyrolysis of DNA Test Results 1. Inorganic Phosphate 2. Purines 3. Deoxyribose
A: Hydrolysis of DNA will cause the DNA backbone to break down and ultimately give the deoxyribose…
Q: Both protein and DNA are run together in an isoelectric focusing (IEF) electrophoresis using the…
A: Isoelectric focusing electrophoresis is a technique which is used for separation of amphoteric…
Q: Solution B that was used during the plasmid isolation contains 0.2 M NaOH (see practical manual).…
A: INTRODUCTION E. coli E. coli are small gram negative motile bacilli which mainly cause urinary…
Q: Which of these is not considered a method of DNA extraction? organic filtration differential…
A: DNA (deoxyribonucleic acid) extraction is a procedure to isolate DNA from the cells. It is a method…
Q: Fluorescence confocal microscopy (FCM) - STK38 monoclonal antibody (M01), clone 2G8- 1F3. 200 μm a.…
A: The STK38 Monoclonal Antibody from LifeSpan BioSciences is a Mouse Monoclonal antibody. This…
Q: Ethidium bromide used to be a common stain for detecting DNA/ RNA on agarose gel. (i) Describc how…
A: During the extraction of DNA fragments via agarose gel electrophoresis, EtBr or ethidium bromide is…
Q: What are the roles of the following reagents in DNA extraction? a. Ethanol b. NaCl c. SDS d. TE…
A: A) The initial role of the ethanol and monovalent cations is to remove the solvation shell…
Q: DNA concentration is 3.75 ng/ ul = the dilution factor is 1:______ we add____ uL dna extract to ____…
A: DNA shows an absorbance maxima at 260 nm and the absorbance is proportional to the concentration of…
Q: During nucleic acid hybridization, the probe is labelled O for DNA stability O to increase…
A:
Q: If the purity of DNA sample is below 1.8 A260/A280, where did the protein contamination come from?
A: The absorbance ratio at 260 nm & 280 nm is used to determine the quality of DNA & RNA. A…
Q: DNA concentration in cuvette solution (in microgram per ml) Dilution factor of DNA sample in cuvette…
A: DNA is the nucleic acid and it absorbs the UV light with maximum absorbance at 260 nm wavelength due…
Q: After incubation, the plates appeared as below: Water EvaGreen Ethidium Bromide SYBR Green 1
A: Ames assay is a method is used to determine whether a chemical can function as mutagen or not by…
Q: One micromole of 48-nucleotide(nt) DNA was synthesized via solid phase synthesis. This sample…
A: DNA or deoxyribonucleic acid is a polynucleotide chain made of monomeric units of nucleic acids.…
Q: Short tandem repeats (STR) profiling is based on
A: Small tandem repeats (STRs) are short repeating DNA sequences (2–6 bp) that make up roughly 3% of…
Q: a. Preparing/determining the concentration of the agarose gel- b. Loading the samples- c. Choosing…
A: a. Preparing/determining the concentration of the agarose gel- The more the concentration of the…
Q: Gel electrophoresis separates molecules based on size O charge O weight What is the restriction cut…
A: Gel electrophoresis is a technique by means of which the DNA molecules are separated on the basis of…
Q: The solid matrix that is usually used to attach the target DNA in southern blotting technique is…
A: Blotting is utilized in atomic science for the distinguishing proof of proteins and nucleic acids…
Q: During nucleic acid hybridization, the probe is labelled Question 1 options: for DNA stability to…
A: Since you have asked multiple question, we will solve the first question for you. If you want any…
Q: Which plate could represent the results of the transformation experiment positive control plated on…
A: Transformation can be defined as the process in which the plasmid of the bacteria is replicated. It…
Q: Complete this Master Mix table for 8 DNA samples, a positive control, negative control, and an extra…
A: Volume per tube (uL) number of reactions volume in master mix (uL) PCR buffer 1.2 11 13.2 dNTP…
Q: Why differential centrifugation can cause contamination in pellet?
A: Differential centrifugation is a typical process used in microbiology and cytology to separate…
Q: Will reveal RFLP polymorphism between the DNA samples after probe detection/visualization; and
A: Restriction Fragment Length Polymorphism (RFLP) is a distinction in homologous DNA sequences that…
Q: Using a 1 cm cuvette, the absorbance at 260nm of your double-stranded DNA sample is 0.15. What is…
A: The concentration of DNA in a sample can be estimated by different methods - absorbance (optical…
Q: During DNA profiling, DNA nucleotides hybridized with probe can be detected througha)…
A: DNA profiling is performed at many loci to be able to tell the genetic difference between different…
Q: n assessing extracted DNA quality, what does A260 indicate? carry-over salts nitrogenous bases…
A: DNA isolation and purification involve disruption and lysis of the cells followed by removal of…
Q: Nucleic Acid Hybridization Technique Northern Blot Western Blot Transblot Molecules involved (DNA,…
A: Nucleic acid hybridization technique Northern blot Western blot Transblot…
Q: In which reagent is extracted DNA suspended to put it in solution? O Sodium dodecyl sulfate (SDS)…
A: Deoxyribonucleic acid is a polymer made up of two polynucleotide chains that coil around one another…
Q: The concentration of your RNA solution is 1500 ng/μl. How much RNA solution do you need to use when…
A: The concentration of RNA in solution can be determined by measuring absorbance at 360 nm.
Q: You have just transformed 1 ul (100 pg/ul) of control pWasabi (plasmid) DNA into 50 ul of E. coli…
A:
Q: DNA could be denatured without maximum damages a. NaCl b. High heat c. NaOH d. Na-acetate
A: The process of separation of double stranded DNA (dsDNA) into single strands by physical or chemical…
Q: Explain 2 possible consequences of accidentally puncturing one of the wells in the agarose gel in an…
A: The chromosomes in our cell cores comprise of huge strands of DNA. These strands are awkward when…
Q: why does a higher agarose concentration render better resolution/separation of smaller DNA…
A: Electrophoresis is an essential lab procedure at the molecular level that uses an electric field to…
Q: Discuss the use of gel electrophoresis for the separation of macromolecules (DNA, RNA and protein)…
A: In biotechnology the gel electrophoresis is routinely used technique that is used for the separation…
Q: proteins are denatured and eliminated during the DNA extraction process. how can we determine the…
A: Deoxyribonucleic acid (DNA) is the hereditary unit of life, which carries the genetic information in…
Q: What are the importance of accurate wavelength measurement in purified DNA sample? Explain why…
A: DNA can be extracted by using different protocols. Protocols of DNA isolation are modified according…
Q: For a DNA sample, the OD value at 260nm = 0.125, and the OD value at 280nm 0.065. %3D Purity of DNA…
A: Purity of a DNA sample is defined as the ratio of absorbance at 260 nm and 280 nm. If the ratio…
Q: Quantity of total DNA in a 5ul sample: Absorbance at 260nm 0.002 and at 280nm = 0.0011
A: The quantification of nucleic acid is a process of determination of the average concentration of DNA…
Q: The DNA fragments separated on an agarose gel can be visualised after staining with what?
A: Gel electrophoresis is a technique used to separate the DNA, RNA, and protein molecules on the basis…
Q: give the significance/role/effect of the reagent/condition in the isolation or analysis of a…
A: DNA isolation is a process of isolation of DNA from biological sample like body fluid, tissue, etc.…
Q: Process of Dpph free radical scavenging assay
A: DPPH free radical scavenging assay is a fast, basic, reasonable and broadly utilized technique to…
Q: 4. is added during the precipitation step of DNA extraction to neutralize charge on nucleic acid…
A: DNA is a hydrophilic substance in its natural state. Hydrophilic solutions dissolve quickly in water…
Q: Double stranded DNA is denatured in the presence of high heat low heat high pressure acidic…
A: According to Bartleby guidelines , we are required to attempt first question in case of multiple…
Q: Which of the following methods for determining protein concentration is most commonly used? O Lowry…
A: “Since you have asked multiple questions, we will solve the first question for you. If you want any…
Q: Why the protien contamination happens during DNA isolation how the isolated DNA get contaminated by…
A: During the cycle of DNA isolation, the DNA test more often get contaminated with RNA and protein. To…
Q: In which reagent is extracted DNA suspended to put it in solution? O Sodium dodecyl sulfate (SDS) O…
A: Introduction DNA extraction is a method to isolate DNA in a biological sample, which is done by…
Step by step
Solved in 3 steps
- 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 IWhat volume of 4X loading buffer must be added to 21 micro L of DNA in a technique for DNA sample preparation for agarose gel electrophoresis to generate a 1X buffer solution?Estimate the viscosith of 1.0 vol % agarose gel solution if it took 58 minutes for 1500 base pairs long DNA molecule to migrate 11cm during gel electrophoresis under 110V. Assume appropriate numerical values for the missing parameters such as the charge and diameter of DNA molecules.
- 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?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.…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…
- 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.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
- what type of gel(in terms of material) can be more suitable for the electrophoresis of 500-1000bp DNA fragment and why?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,…Agarose gel electrophoresis is a technique used to separate DNA fragments on an agarose matrix. The DNA is then viewed by staining under UV light. (i) Explain the principle for the migration of DNA molecules in an electric field. DNA loading dye is used to prepare DNA markers and samples for loading on the agarose gel. State the function of bromophenol blue in the loading dye. (ii)