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

Biochemistry
6th Edition
ISBN:9781305577206
Author:Reginald H. Garrett, Charles M. Grisham
Publisher:Reginald H. Garrett, Charles M. Grisham
Chapter28: Dna Metabolism: Replication, Recombination, And Repair
Section: Chapter Questions
Problem 17P
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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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