C. How much does DNA of a WBC weigh? (Note that WBCs are diploid.) d. How much DNA can you collect from 10 mL of whole blood? (Assume that there is no loss during DNA extraction.)
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- 1. You are given a tube containing 1.5 ml of a DNA solution. The concentration of the DNA solution is 1.2 µg/ul. a. What is the concentration of the DNA solution in mg/ml? b. How much total DNA is present in the DNA solution? Give the mass of DNA in both ug and mg.2. Below is an example of an image of genomic DNA extracted from whole blood analysed by agarose gel electrophoresis (lanes 4-16). M1 and M2 are molecular weight markers. Using this image as reference, discuss how one would determine the integrity of a genomic DNA sample after extraction, and what one would expect to observe. Critically evaluate the samples analysed here. 23K M1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 M2 bp 2K bp Figure 1: Genomic DNA pattern following 1% agarose gel electrophoresis. 1.5K 500 1002. Suppose you prepared the following six test tubes as indicated in the table below. You divide the material in each tube appropriately into two separate tubes, and pro- ceed to run an orcinol and a diphenylamine test. Tabulate and explain the results you would expect to obtain from the Six orcinol tests and the SIX diphenylamine tests. Reagent DNA (m1) RNA (ml) Water (m1) 0.5 N KOH (ml) 10% TCA (m1) 1 2 ININ 2 2 2 2 1 Test Tube Number 3 4 2 2 2 2 5 2 NIN 2 6 2 2
- 4. You isolate DNA from 1 ml of a suspension of E. coli cells using the procedure outlined in your lab handout. After completing the DNA isolation procedure, the total volume of the DNA solution is 700 µul. You transfer 80 µl of this DNA solution into 720 µl of TE buffer to create a diluted DNA solution. Using a spectrophotometer, you determine that the concentration of the diluted DNA solution is 440 µg/ml. a. How much did you dilute the original DNA solution? Express the dilution as a ratio. b. What is the concentration of the original DNA solution? c. How much total DNA did you isolate from the suspension of E. coli cells? d. How much total DNA was present in the diluted DNA solution?3. You isolate DNA from 1 ml of a suspension of E. coli cells using the procedure outlined in your lab handout. After completing the DNA isolation procedure, the total volume of the DNA solution is 600 ul. You transfer 10 µl of this DNA solution into 490 µl of TE buffer to create a diluted DNA solution. Using a spectrophotometer, you determine that the concentration of the diluted DNA solution is 245 ug/ml. a. How much did you dilute the original DNA solution? Express the dilution as a ratio. b. What is the concentration of the original DNA solution? c. How much total DNA did you isolate from the suspension of E. coli cells?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…
- When Griffith injected mice with a combination of live rough-strain and heat-killed smooth-strain pneumococci, he discovered that (a) the mice were unharmed (b) the dead mice contained living rough-strain bacteria (c) the dead mice contained living smooth-strain bacteria (d) DNA had been transferred from the smooth-strain bacteria to the mice (e) DNA had been transferred from the rough-strain bacteria to the smooth-strain bacteria1. Answer the following A. What is the relationship between the DNA molecular weight and the distance travelled by DNA fragments from the well? Create a line diagram with DNA molecular weight as the X axis and distance travelled as the Y axis. B. How is the DNA charge and molecular weight considered in agarose gel electrophoresis?4. After reading the lab for next week, use the following simulated DNA gel to answer the questions. a. Label the bands of the two-log ladder with the lowest band as the lowest number of kilobases (far left column or lane) b. Draw an arrow indicating the direction the DNA fragments travelled c. Estimate the number of kilobases in the unknown DNA in lanes 7 and 8 MW 1 2 3 4 5 6 7 8
- 1. Compute the volumes needed to complete this table. Stock Final Volume for Volume for 5x reaction Reaction Component Concentration Concentration 1x reaction sddH,0 up to 25 ul up to 25 ul Buffer (ViBuffer A) 10X 1X MgCl, 50 mM 3 mM DNTP 2 mM 100 uM forward primer 10 uM 0.2 uM reverse primer 10 uM 0.2 uM Taq polymerase 1 U/ ul 1 U/ 25 ul DNA template 1-2 ng 1 ul *5X reactions = 2 samples, 1 NTC, 1 positive control, 1 pipetting allowanceAnswer each of the following correctly. Designer Genes Work (This is all about Applications of Recombinant DNA). 1. How does DNA Replicate?(1-3 sentences only) 2. What is Genetically Modified Organism (GMO)?(1-4 sentences only) 3. Illustrate your own Designer genes using this information:The Arctic apple is a fruit engineered to resist browning after being cut. Currently they are only available in the US – in golden, fuji and gala varieties – where they have been given Food and Drug Administration approval. If approved in Europe, they would have to be labelled as genetically modified. The manufacturers claim the main benefit is to help cut down on food waste. And based on the following: 1. Identify a special trait. 2. Identify a source organism. 3. Identify a target organism 4. Identify the modified/added trait. Example: Hot Tomato > Chili > Tomato > Spicy Tomato It was reported this week that Brazilian scientists are hoping to create spicy tomatoes using Crispr…1c) T. aquaticus genomic DNA is 34.3% guanosine nucleotides. What fraction of the DNA is adenosine nucleotides? 1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 Fraction of DNA