For human genomic DNA what is the expected fragment size for high molecular weight DNA extracts? What is an important component of gel electrophoresis which is omitted from this result, why would it be important to include ?
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- For human genomic DNA what is the expected fragment size for high molecular weight DNA extracts?
- What is an important component of gel electrophoresis which is omitted from this result, why would it be important to include ?
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- 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…Use the figure of the gel below to answer the following questions. Controls 1 2 3 Experimentals 4 5 6 7 8 8. Identify which well contains the DNA ladder-Helps identify unknown DNA to which individual. 9. Identify which well contains a smear-Contamination of the well or degrading the DNA 10. Identify which well contains primer dimer. 11. Identify which well shows results similar to the positive control.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
- 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…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…Figure 4 shows the results from Nanodrop quantification of DNA sample. Sarmple ID E Coli (K-12) Pedestal 14 Турe DNA 50 00 Conc. 843.6 ng/ul A260 (10 mm path) A280 (10 mm path) 260 / 280 2.11 260 / 230 2.24 O Baseline correcton 340 nm Wavelength (nm) Figure 4 (i) Interpret the Nanodrop result above. (ii) How to improve the quality of the extracted DNA in order to obtain a better result? (ii) Calculate the absorbance of DNA at 260nm. 10mm Absorbance
- Question Image hown bele nallest? Q. A gel from gel electrophoresis is shown below. Which DNA fragment is the smallest? B. Direction of TravelFor a DNA sample, the OD value at 260nm = 0.125, and the OD value at 280nm 0.065. Purity of DNA - OD 260/280 %3D Concentration of DNA = Abs x 50 µg/ml x dilution factor (100) 0.125 x 50 µg/ml x 100 = Edit View Insert Format Tools Table 12pt v Paragraph v BIUA 2 T'vComplete this Master Mix table for 8 DNA samples, a positive control, negative control, and an extra reaction for pipetting error. Show your work. Master Mix Conc. Of Total μL µL/Rxn (total per tube) Final Number of Stock Conc. Reactions needed for solution master mix PCR buffer 50X 1X Water DNTP mix 100 mM 200 µM MgCl2 Forward 24 mM 2.5 mM 2 μΜ 0.1 μΜ primer Reverse 2 μΜ 0.1 μΜ primer Taq polymerase DNA 5 U/uL 0.03 U/ µL 1 μL 60 μL Total volume of the entire reaction (µL)
- An AGE run was set at 100V for 30 min, where 3 ul of the ladder (100 ng/ul) was loaded into the gel, while 10 ul of the DNA samples plus an appropriate amount of 6x loading buffer were added into the gel. Shown below is the AGE profile and the needed marker details where M is the MW marker while numbers 1-4 indicate the lanes with the DNA samples: M 2 3 4 ng/0.5 µg bp 5000 20 4000 40 3000 20 2000 20 1000 60 500 100 Compare the bands in the Lanes 1,2,3,4 with the nearest DNA standard by determining the DNA standard which is nearest in size to the samples in Lanes 1,2,3,4 DNA standard (choices: 100, 500, 1000, 2000, 3000, 4000, or 5000 bp) DNA sample Lane 1 bottom band Lane 1 top band Lane 2 band Lane 3 band Lane 4 band 20 20An AGE run was set at 100V for 30 min, where 3 ul of the ladder (100 ng/ul) was loaded into the gel, while 10 ul of the DNA samples plus an appropriate amount of 6x loading buffer were added into the gel. Shown below is the AGE profile and the needed marker details where M is the MW marker while numbers 1-4 indicate the lanes with the DNA samples: м 1 M 2 3 4 ng/0.5 µg bp 20 5000 20 4000 40 3000 20 2000 20 1000 60 500 20 100 Determine the amount of the total DNA ladder added in the gel in terms of ug5. Show the separation pattern of the following DNA molecules on an agarose gel electrophoresis. 5 kbp 5 kbp 5 kb 5 kbp ----