DNA Extraction by Alkaline LysisProcedure:1. Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for 20s topellet. 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 difficultto resuspend cells.2 Resuspend pellet in 100pul GTE solution and let sit 5 min at room temperature. Be surecells are completely resuspended.3. Add 200ul NaOH/SDS solution, mix by tapping tube with finger, and place on ice for 5min.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 DNA6. Transfer 0.4 ml supernatant to a fresh tube, mix it with 0.8 ml of 95% ethanol or 0.4ml 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 RNA.8.Remove supernatant, wash the pellet with 70% ethanol, and dry pellet under vacuum.9. Resuspend the pellet in 30ul TE buffer and store at 4'C. Contaminating RNA mayinterfere with detection of DNA fragments on the agarose gel; it can be destroyed byadding Iul of a 10 mg ml RNase solution (DNase-free).Isolation of DNA from BananaProcedure:1) The precipitating agent was prepared by adding 1 tablespoon of salt and 5 tablespoonsofiquid soap. Then, 100mL of hot water was mixed and stirred evenly.2) A banana was peeled and chopped into small pieces, then it was placed in a bowlwithsufficient water and then mixed with the precipitating agent.3) The banana was then mashed with a fork.4) The mashed banana was then filtered through gauze with a strainer and thefiltrate iscollected in a transparent container.5) The methylated spinit was then taken out from the fridge and mixed with the filtrategently, and the sample is observed for changes for 2-3 minutes.6) The methylated spirit was then taken out from the fridge and mixed with the filtrategently, and the sample is observed for changes for 2-3 mimnutes.Compare and contrast the difference between this two protocol (DNA Extraction by AlkalineLysis and Isolation of DNA from Banana)

To isolate plasmid DNA, cells are cracked open and a mini preparation should be performed, trying…

Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for pellet. Remove the supernatant completely with a Pasteur pipet or a plastic pipet The spins can be performed at C or at room temperature. Longer spins make it a to resuspend cells. Resuspend pellet in 100pl GTE solution and let sit 5 min at room temperature. A cells are completely resuspended. Add 200al NaOH/SDS solution, mix by tapping tube with finger, and place on ice min. Add 150ul potassium acetate solution and vortex at maximum speed for 2s t Place on ice for 5-15 min. Be sure mixing is complete. Spin 3 min at 12,000 rpm to pellet cell debris and chromosomal DNA

Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for pellet. Remove the supernatant completely with a Pasteur pipet or a plastic pipet The spins can be performed at C or at room temperature. Longer spins make it a to resuspend cells. Resuspend pellet in 100pl GTE solution and let sit 5 min at room temperature. A cells are completely resuspended. Add 200al NaOH/SDS solution, mix by tapping tube with finger, and place on ice min. Add 150ul potassium acetate solution and vortex at maximum speed for 2s t Place on ice for 5-15 min. Be sure mixing is complete. Spin 3 min at 12,000 rpm to pellet cell debris and chromosomal DNA

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 2DAA: HersheyChase Experiments The graph shown in FIGURE 8.5 is reproduced from an original 1952...

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Drag and drop the appropriate text in the gaps below. Firstly, the plasmid DNA in the E. coli is propagated through Then the bacterial cells are pelleted by centrifugation at 10,000 RPM. The media supernatant is removed and the bacterial cells are The bacterial cells are then lysed via |. Contaminating macromolecules such as protein and chromosomal DNA is removed by Overnight incubation of the bacterial culture washed in cell resuspension buffer addition of lysis buffer addition of neutralisation buffer washing of the spin column separation via agarose gel electrophoresis
biotechnology lab class :1. The bacterial streaking on antibiotic containing agar plates has been completed and the plates will be available for you to inspect in the lab class.2. You will complete a restriction enzyme digest on the four available plasmid stocks and then examine the products by agarose gel electrophoresis. Photographs of the gel are attached to identify the plasmid present in each analysed sample.Now consider the following questions :- For each of the four plasmids used in the practical, identify the size (in kb) of the linear DNA fragment(s) that you would expect to obtain if the EcoRI digest is complete.- Using the provided negative image of the gel, which shows the bands present in the ND and D samples for each plasmid, identify the plasmid that was present in each of the four plasmid stock.- Explain how you were able to identify plasmid in each sample using the gel, considering how linear DNA fragments of different length are separated by agarose gel…
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?
In which reagent is extracted DNA suspended to put it in solution? O Sodium dodecyl sulfate (SDS) Sodium citrate saline buffer Cold absolute alcohol OH2SO4 O HCI
Which of the following statements is TRUE for DNA extraction? DNA yield and purity are inversely proportional. The same protocol can be used for isolating bacterial gDNA and pDNA. OCTAB extraction is only effective for plant cells. extraction is safe routine Phenol-chloroform protocol.
Which of the following gels is the preferred choice for separation of DNA fragments? Agarose Polyacrylamide SDS PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis) Agar
DNA Cheek Cells Lab Identify the component of the solution which is the extracted DNA
A amp PBR322 4301 fot B Clear Zones Figure 2 The postgraduate student, Demika, inserted her gene of interest into the plasmid, pBR322, before transformation into the competent host cell using heat shock method. After that she cultured the cells on the Ampicillin agar plate before replica plating the colonies onto another Ampicillin (A) and Tetracycline (B) agar plates shown in Figure 2. (1) Referring to the vector pBR322 in Figure 2, which recognition site was cleaved to insert the gene of interest? Based on the observation above, can you identify which colonies are carrying positive mcombinants of BR322? Explain your selection.
Which of the given statements is correct in the context of visualizing DNA molecules separated by agarose gel electrophoresis? DNA can be seen in visible light DNA can be seen without staining in visible light Ethidium bromide-stained DNA can be seen in visible light Ethidium bromide-stained DNA can be seen under exposure to UV light Please explain the answer properly and kindly relate to the NCERT Bio XII chapter 11.
Examine the pGLO plasmid DNA solution with the UV lamp. Note your observations. Using a micropipettor, withdraw 10 ul of plasmid and mix it into the cell suspension of the +PGLO tube by pipetting up and down three times. Close the tube and return it to the rack on ice. Also close the -PGLO tube. Do not add plasmid DNA to the -pGLO tube. Why not?
The rate of migration of DNA within an agarose gel in the gel electrophoresis technique is primarily based on: O The number of the DNA fragments. The size of the DNA fragments. The concentration of the DNA None
Complete 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)