Homework Help is Here – Start Your Trial Now!
Science
Biology
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

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
See similar textbooks
icon
Related questions
Question
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 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 may interfere with detection of DNA fragments on the agarose gel; it can be destroyed by adding Iul of a 10 mg ml RNase solution (DNase-free). Isolation of DNA from Banana Procedure: 1) The precipitating agent was prepared by adding 1 tablespoon of salt and 5 tablespoons ofiquid 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 bowl withsufficient 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 the filtrate iscollected in a transparent container. 5) The methylated spinit was then taken out from the fridge and mixed with the filtrate gently, 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 filtrate gently, and the sample is observed for changes for 2-3 mimnutes. Compare and contrast the difference between this two protocol (DNA Extraction by Alkaline Lysis and Isolation of DNA from Banana)
Transcribed Image Text: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 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 may interfere with detection of DNA fragments on the agarose gel; it can be destroyed by adding Iul of a 10 mg ml RNase solution (DNase-free). Isolation of DNA from Banana Procedure: 1) The precipitating agent was prepared by adding 1 tablespoon of salt and 5 tablespoons ofiquid 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 bowl withsufficient 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 the filtrate iscollected in a transparent container. 5) The methylated spinit was then taken out from the fridge and mixed with the filtrate gently, 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 filtrate gently, and the sample is observed for changes for 2-3 mimnutes. Compare and contrast the difference between this two protocol (DNA Extraction by Alkaline Lysis and Isolation of DNA from Banana)
Expert Solution
trending now

Trending now

This is a popular solution!

steps

Step by step

Solved in 3 steps

SEE SOLUTIONCheck out a sample Q&A here
Blurred answer
Knowledge Booster
Microbial isolation
Learn more about
Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.
Similar questions
  • SEE MORE QUESTIONS
Recommended textbooks for you
Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
Biology
ISBN:
9780134580999
Author:
Elaine N. Marieb, Katja N. Hoehn
Publisher:
PEARSON
Biology 2e
Biology 2e
Biology
ISBN:
9781947172517
Author:
Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:
OpenStax
Anatomy & Physiology
Anatomy & Physiology
Biology
ISBN:
9781259398629
Author:
McKinley, Michael P., O'loughlin, Valerie Dean, Bidle, Theresa Stouter
Publisher:
Mcgraw Hill Education,
Molecular Biology of the Cell (Sixth Edition)
Molecular Biology of the Cell (Sixth Edition)
Biology
ISBN:
9780815344322
Author:
Bruce Alberts, Alexander D. Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter Walter
Publisher:
W. W. Norton & Company
Laboratory Manual For Human Anatomy & Physiology
Laboratory Manual For Human Anatomy & Physiology
Biology
ISBN:
9781260159363
Author:
Martin, Terry R., Prentice-craver, Cynthia
Publisher:
McGraw-Hill Publishing Co.
Inquiry Into Life (16th Edition)
Inquiry Into Life (16th Edition)
Biology
ISBN:
9781260231700
Author:
Sylvia S. Mader, Michael Windelspecht
Publisher:
McGraw Hill Education
SEE MORE TEXTBOOKS