pOpen-K12polLF (exo-)
(Plasmid #165522)

• Purpose: DNA pol fragment used in flourescent labelling for microarray, dA and dT tailing, and ligating DNA adapters to DNA fragments.
• Depositing Labs: Drew Endy, Jennifer Molloy, FreeGenes Project
• Publication: Open Enzyme Collection (unpublished)

Backbone

• Vector backbone: pBuild
• Vector type: Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: DNA Polymerase I, Large (Klenow) Fragment (3'-5' exo-)
• Species: Other

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AATG (unknown if destroyed)
• 3′ cloning site: GCTT (unknown if destroyed)
• 5′ sequencing primer: M13 forward
• 3′ sequencing primer: M13 reverse

Terms and Licenses

• Academic/Nonprofit Terms:
  • OpenMTA
• Industry Terms:
  • OpenMTA

Depositor Comments

DNA polymerase fragment used in fluorescent labelling for microarray, dA and dT tailing, and important in the process of ligating DNA adapters to DNA fragments. Frequently used in preparing DNA libraries for Next-Gen sequencing.The exo-Klenow polymerase is a mutant polymerase from the Klenow DNA polymerase large fragment that lacks 5'-3' polymerase activity and 3'-5' exonuclease activity.

The plasmids in the Open Enzyme collection contain genetic parts and are not functional by themselves. Researchers can clone these enzymes into an expression vector of their choice and then transform into E. coli bacteria for expression and purification. Additional information and protocols can be found at https://stanford.freegenes.org/