pOpen-BstpolLF
(Plasmid #165553)

• Purpose: Fragment retains 5'-3' polymerase activity from full length Bst DNA Polymerase, while lacking 5'-3' exonuclease activity. Suitable for applications requiring thermophilic strand displacement.
• 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• Publication: Open Enzyme Collection (unpublished)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: Bst DNA Polymerase, Large Fragment
• 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

Fragment retains 5'-3' polymerase activity from full length Bst DNA Polymerase, while lacking 5'-3' exonuclease activity. Suitable for applications requiring thermophilic strand displacement. Additionally used for Loop-Mediated Isothermal Amplification (LAMP), DNA sequencing through high GC regions, rapid sequencing from nanogram amounts of DNA template. Can be used in the standard Sanger one step or two step protocol which separates the labelling reaction from the elongation termination reaction. The enzyme can be used in double stranded sequencing.

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/