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- 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: 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 ugWhy 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…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
- An AGE run was set at 100V for 30 min. The 3 ul of the ladder 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. Find the amount of the 6x loading buffer added to 10 ul DNA samples in order to make the samples sink in the gel? Tip. From 6x final conc of the loading buffer should be 1x and answer shoud be in ulAn AGE run was set at 100V for 30 min. The 3 ul of the ladder 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. Find the amount of the 6x loading buffer added to 10 ul DNA samples in order to make the samples sink in the gel? Tip. From 6x final conc should be 1x and answer shoud be in ulFor 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'v
- 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?Given the electrophoresis profile of a Sanger sequencing result, what was the sequence of the original DNA sample used for sequencing? ddATP ddGTP ddCTP ddTTP - GGTTACC B CCATTGG CCAATGG GGTAACC | | | |Aa Po AaBbCcDc AaBbCcDdEe AaBbCcDdE AaBbCcDdE AaBbCcDdE BIUVab x, x A ev A 三==|這、|の Caption Emphasis Heading 1 Heading 3 Heading 4 Style Pane (a) Complete the table, using a letter D, R or B, to show whether each statement applies to: Fo DNA only (D) TA2G – Completed forms must be available for Open Awards external moderation purposes. Page 6 of 15 > T | RNA only (R) Both DNA and RNA (B) Statement D or R or B Contains thymine Contains ribose Consists of two chains connected to each other with hydrogen bonds Has a sugar-phosphate backbone Has four different nitrogenous bases Contains a pentose sugar Is found in the nucleus and cytoplasm English (United Kingdom) Focus 目民 2490 words +1
- 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 choiceComplete 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)If the DNA was replicated using the dispersive model, what would you have expected to observe in the generation 2 sample? Select an answer and submit. For keyboard navigation, use the up/down arrow keys to select an answer. a b с d e Question 6 Homework Answered 6.0 g 3 distinct bands (one 15N, one 14/15N, and one 14N) 2 distinct bands (one 15N and one 14N) 2 distinct bands (one 15N and one 14/15 hybrid) Only one distinct band (15N) f Only one distinct band (14N) Only one distinct band (14/15 hybrid) No bands