You have run a gel with 5 uL of amplified DNA in addition to 1 uL of 6X loading dye. Comparing the brightness of this band to your marker, you estimated that your amplified DNA is approximately 62 ng. What was the concentration of your amplified DNA that was loaded? You do not need to account for the volume of loading dye in your calculations.
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- What is the concentration of a DNA solution that absorbs 0.812 and 0.463 at 260 and 280 nm, respectively? Is the DNA solution considered to be good quality? Why or why not?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…How would you estimate the size of the unknown DNA fragement just by looking at the gel?
- A facility says they need 15 μL of a 40 ng/μL solution of plasmid DNA for sequencing. The typical yield for a DNA miniprep 5 μg eluted in 50 μL of solution. What do you need to do (for example dilution) to send the appropriate amount to the facility? Show all math work with an explanation.A sample of DNA with the sequence 5'- CTC GAG CGA AGC TCA ACC-3' he was obtained as a dry solid. The sample was dissolved in 1000 µl of deionized water ane mixed well. Ten microliters of the dissolved sample was then transferred to a new sampie tube and mixed with 990 ul of water, The dilute sample had A260 = 0.156. What was the concentration of the original sample (the solid dissolved in 1000 ul of deionized water)Please answer You have received a dehydrated sample of DNA primer at a concentration of 19.9 microM. what volume (in microlitres) of buffer would you add to achieve a solution of 100nM of this primer? Show workings.
- Visually compare the intensity of your team’s Plant-DNA sample with the intensity of the 3 kb band on the DNA ladder (see figure posted below). For example, if your Plant- DNA sample closely matches the 3 Kb band, this represents approximately 125 ng of total DNA. Divide the approximate mass (ng) of DNA in your sample by the volume loaded in the gel minus the loading dye added (i.e., 12 – 2 = 10 μL, since you loaded 12 μL from a 24 μL sample that contained 20 μL DNA prep + 4 μL loading dye. What is the DNA concentration of your sample? ______ng/μLYou extract DNA from 200, You pipette 200 microlitres of this extraction sample into a 3-ml cuvette and make up to 3.0 ml using buffer. The absorbance at 260 nm (A260) of the solution in the cuvette is 0.2 (Remember: an A 260 of 1.0 corresponds to a DNA concentration in the cuvette solution of 50 micrograms per ml). Calculate the total amount of DNA in 100g wheat germ, assuming that the extraction efficiency of DNA from wheat germ during your lab protocol is 40%. Please fill out each of the boxes in the Table below What to Calculate Calculation / Result Marks (x / 100) 12/100 DNA concentration in cuvette solution (in microgram per ml) Dilution factor of DNA sample in cuvette 12/100 12/100 Total amount (microgram) of DNA in the entire extraction sample Total amount (milligram) of DNA in 100g wheat germ 24/100 assuming an extraction efficiency of 40%You have 10 ul of DNA you want to cut with HindIII, and then run a gel, in order to cut out certain bands and purify the DNA from them. Since the DNA is pretty concentrated, you are going to cut it in a 50 ul total volume and then run it in 2 adjacent lanes of a gel, to avoid overloading the lanes. What are all the ingredients you need to put into a tube, how much of each, and what is the temperature ordinarily used? (Amount of HindIII might vary, but be safe and put in twice as much as you might normally use to cut less DNA)
- You extract DNA from 200 milligram of wheat germ. Your total volume of DNA extraction sample is 500 microlitres. You pipette 200 microlitres of this extraction sample into a 3-ml cuvette and make up to 3.0 ml using buffer. The absorbance at 260 nm (Az60) of the solution in the cuvette is 0.2 (Remember: an Az60 of 1.0 corresponds to a DNA concentration in the cuvette solution of 50 micrograms per ml). Calculate the total amount of DNA in 100g wheat germ, assuming that the extraction efficiency of DNA from wheat germ during your lab protocol is 40%. Please fill out cach of the boxes in the Table below What to Calculate | Calculation / Result DNA concentration in cuvette solution (in microgram per ml) Dilution factor of DNA sample in cuvette Total amount (microgram) of DNA in the entire extraction sample Total amount (milligram) of DNA in 100g wheat germ assuming an extraction efficiency of 40% In your extraction of DNA from wheat germ, would you expect that nuclear DNA is the only source…You extract DNA from 200 milligram of wheat germ. Your total volume of DNA extraction sample is 500 microlitres. You pipette 200 microlitres of this extraction sample into a 3-ml cuvette and make up to 3.0 ml using buffer. The absorbance at 260 nm (A26o) of the solution in the cuvette is 0.2 (Remember: an A260 of 1.0 corresponds to a DNA concentration in the cuvette solution of 50 micrograms per ml). Calculate the total amount of DNA in 100g wheat germ, assuming that the extraction efficiency of DNA from wheat germ during your lab protocol is 40%. Please fill out each of the boxes in the Table belowBoth 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.