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- sampies B. In this picture of gel electrophoresis of DNA, which sample contains the longest/largest piece of DNA? O A. A O B. BMake your DNA extraction buffer. Add 2 teaspoons of detergent, 1 teaspoon of salt, and ½ cup of water to one of the plastic cups. Stir well. 4 Open the bag of fruit and add 2 teaspoons of your DNA extraction buffer. What are cell membranes made of? Which parts are hydrophobic and which are hydrophilic? What does soap do to cell membranes? Why would this be helpful in extracting DNA? What function does the salt perform in the solution?BIOTECHNOLGY Date: Name: Instructor: Section/Group:. POST-LAB QUESTIONS 1. In one or two sentences, summarize the technique of gel electrophoresis. 2. How does the process of gel electrophoresis separate DNA fragments? 3. Why is the fact that DNA has a negative charge so important in the gel electrophoresis process? Biotechnology 165
- Indicate true or false for the following statements The glycerol used in the DNA loading dye allows DNA to be visualized under UV light. The DNA Ladder used for agarose gel electrophores can be used to estimate fragment size and DNA concentration. During gel electrophoresis a DNA smear may indicate that DNase was still present in the sample.6. B. samples A and B in the tubes provided. Tube A B Using the spectrophotometer, perform DNA separation purity analysis of 260:280 ratio Mass of empty tube DI Water 2.80 Which of the samples (Tube A or B) represent values consistent with DNA contaminated with residual substances? Explain your rationale. Mass of water and tube 1.80 7. A. (p) 3 microcentrifuge tubes were labeled and the empty mass was recorded. DI water was added to each tube as noted below. The final mass of the tubes containing the water was recorded. Which tube represents the most inaccurate micropipetting? Tube 1 .280g 5uL .284g Tube 2 .280g 50uL .318g Tube 3 .280g 500uL .780gtime and read each question carefully. This is an independent test Question 1 1. If 30% of DNA is Adenine, what percent is Thymine? O 30% O 20% O 40%
- 1. corrected DNA sequences 2.mRNA sequence 3. Amino acid sequence and description 4. Drawing of the ice cream with the labelDNA 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…Procedure: 1. Using the DNA provided transcribe DNA into mRNA. 2. Use the mRNA strand you created and break it up into codons. 3. Plug the codons into the amino acid chart to determine the correct amino acid needed to build that protein. 4. Identify the protein you made by comparing the sequence to the pictures 5. Answer the questions for each protein molecule you build before moving on to the next. Protein 1: DNA A AGACCGTATAC mRNA Amino Acid Sequence 1. Which kind of protein molecule did this gene make? 2 How does this protein help the body maintain homeostasis2
- 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…2. 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 2Samples/Enzyme DNA fragment(s) length 1. None 1500 2. ECORI 700, 800 3. BamHI 300, 1200 4. EcoRI and BamHI 300, 500, 700 А. Using the line below, draw a map showing the locations for each enzyme digestion site. Include distances. _1500 Use paper and draw more lines and try different combinations of cuts until you get the fragments with the right dimensions. Remember that I am asking to have resulting fragments long 300, 500 and 700 bps but they DO NOT have to be in that order.