pOpen-TthpolA
(Plasmid #165533)

• Purpose: DNA Polymerase whose activity is resistant to prolonged incubations at high temperatures (95 degrees C).
• 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• Publication: Open Enzyme Collection (unpublished)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: Tth DNA polymerase
• 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 whose activity is resistant to prolonged incubations at high temperatures (95 degrees C); can therefore be used for PCR amplification of DNA fragments. Also accepts modified deoxyribonucleotides as substrates and can thus be used to label DNA fragments with radiolabeled nucleotides, digoxigenin, or biotin. Intrinsic Mn-dependent RT activity allows the enzyme to efficiently transcribe RNA targets into cDNA. Tth DNA Polymerase has been used for real time PCR.

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/