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for a 5:1 insert: vector ligation reaction, including the volumes of insert and vector you calculated above, and the volumes required for a 20 uL reaction: L

for a 5:1 insert: vector ligation reaction, including the volumes of insert and vector you calculated above, and the volumes required for a 20 uL reaction: L

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,...
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Note that the table provided shows a ligation using a molar ratio of 1:3 vector to insert. Write out the complete recipe for a 5:1 insert: vector ligation reaction, including the volumes of insert and vector you calculated above, and the volumes required for a 20 uL reaction:

  • Ligase buffer 
  • Nuclease-free water 
  • T4 DNA ligase 
COMPONENT T4 DNA Ligase Buffer (10X)* Vector DNA (4 kb) Insert DNA (1 kb) Nuclease-free water T4 DNA Ligase 20 μl REACTION 2 μl 50 ng (0.020 pmol) 37.5 ng (0.060 pmol) to 20 μl 1 μl
Transcribed Image Text:COMPONENT T4 DNA Ligase Buffer (10X)* Vector DNA (4 kb) Insert DNA (1 kb) Nuclease-free water T4 DNA Ligase 20 μl REACTION 2 μl 50 ng (0.020 pmol) 37.5 ng (0.060 pmol) to 20 μl 1 μl
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