Answered: 1. How do you determine the purity of…

1. How do you determine the purity of DNA? If a DNA has 1.8 A260/A280 ratio, what does it mean? What if the DNA has 2.5 A260/A280 ratio?

Biology: The Unity and Diversity of Life (MindTap Course List)

14th Edition

ISBN:9781305073951

Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Chapter8: Dna Structure And Function

Section: Chapter Questions

Problem 2SQ: What are the base-pairing rules for DNA? a. A-G, T-C b. A-C, T-G c. A-T, G-C

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In order to determine the purity of a DNA sample. spectrophotometry can be carried out at calculated, and a number between to measure DNA, and to measure protein. The ratio is then indicates higher purity. 280 nm; 260 nm; A280/A260; 0.5-1 260 nm; 280 nm; A260/A280; 0.5-1 260 nm; 280 nm; A280/A260; 1.5-2 260 nm; 280 nm; A260/A280; 1.5-2
1. Write the complementary base sequence for the matching strand in the following DNA section: -A-G-T-C-C-A-A-T-C–
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?
1. On paper, write out the first 30 bases in the middle of the page in order. You want ensure you get them all in one line, with room to write more information around them. 2. Fill in the complementary base for each of the 30 bases. Label this series of letters “Complementary Bases” on the paper. When you are done, you should have a complete double-helix section of DNA.3. On the opposite side of the original sequence (opposite from where you did the complementary strand), translate the original sequence into mRNA. Label this sequence as mRNA.4. Translate the mRNA strand into amino acids using your codon table. You only need to record the 3-letter code for each amino acid. Label this line as “Amino Acids.” Record this sequence in your lab report (question 8).5. Take a picture of your completed diagram for your lab report. Part 2 – Mutations You will use the same 30-base sequence you started with for this section. 1. Write out the original 30-base DNA sequence on a new paper.2.…
2. Tube A contains 800 µl of a DNA solution that has a concentration of 345 µg/ml. a. You transfer 0.2 ml of DNA solution from Tube A and place it into Tube B. How much total DNA is present in Tube B?
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?
How would the appearance of your DNA gel change if a 0.1% agarose gel slab was used? What about a 7% agarose gel slab? For separating small fragments of DNA, is it better to use a 7% gel or a 0.1% gel? Why?
What are the base-pairing rules for DNA? a. A-G, T-C b. A-C, T-G c. A-T, G-C
Which of the following DNA has a high G-C content? (Tm refers to the melting temperature of the DNA. At the melting temperature, the 2 strands are denatured, i.e. they separate into single strands) DNA Tm A 68 B 49 81 56 Cannot be determined A C E
2. Dr. Kim at Research Center performed shotgun Sanger sequencing on an unknown DNA sample, and obtained the following reads (12 reads). Since the length of each read is quite short, Dr. Kim ran the original sample in a gel electrophoresis, and realized that the original DNA is just 50 base pairs long. (Please note that the resolution of gel electrophoresis is not so good. Thus, we cannot figure out the exact length of DNA using gel electrophoresis in the real world.) 2) AGCCG AGATG GCTG 4) CTGCA AGCCG A 1) САСТС ССAGT GTACC T 3) GGAGT CAATC GC 5) GGCTG TGCTT GG 7) GATGG CTGTG 9) CAGTG TACCT GCA 11) TGCAA GCCGA G 6) CTTGG AGTCA ATCGC 8) САСТС ССAG 10) GCTGT GСТTG G 12) TGCTT GGAGT (a) Find the sequence of the original DNA (reconstruction), and align each read with the reconstructed DNA sequence. (hint: put all reads on a ppt slide, and move them around to find overlaps.) (b) Calculate coverage at each nucleotide position of the reconstructed DNA, i.e., how many reads cover that…
29. You have purified plasmid DNA and would like to determine its concentration. You mix 10 µL of DNA into 990 µl of TE buffer and read the absorbance in a quartz cuvette at 260 nm and 280 nm. The absorbance values are as follows: Azs0 = 1.625 A280 = 0.864 a) Calculate the concentration of the DNA sample b) Given the information above, how could you test the purity of this sample? Is the DNA pure? If not, what are the likely contaminants?
2. Dr. Kim at Ewha Research Center performed shotgun Sanger sequencing on an unknown DNA sample, and obtained the following reads (12 reads). Since the length of each read is quite short, Dr. Kim ran the original sample in a gel electrophoresis, and realized that the original DNA is just 50 base pairs long. (Please note that the resolution of gel electrophoresis is not so good. Thus, we cannot figure out the exact length of DNA using gel electrophoresis in the real world.) 1) САСТС ССAGT GTACC T 3) GGAGT CAАТC GC 5) GGCTG TGCTT GG 7) GATGG CTGTG 9) CAGTG TAССT GCA 11) TGCAA GCCGA G 2) AGCCG AGATG GCTG 4) CTGCA AGCCG A 6) CTTGG AGTCA ATCGC 8) САСТС ССAG 10) GCTGT GСТTG G 12) TGCTT GGAGT (a) Find the sequence of the original DNA (reconstruction), and align each read with the reconstructed DNA sequence. (hint: put all reads on a ppt slide, and move them around to find overlaps.) (b) Calculate coverage at each nucleotide position of the reconstructed DNA, i.e., how many reads cover that…