Manta 1.0 DNA Polymerase

• Thermostable DNA polymerase (Bst)
• Strong strand-displacement activity
• No 5'→3' exonuclease activity and 3'→5' exonuclease activity

Manta 1.0 DNA Polymerase is a Thermostable bacillus (Bst) DNA polymerase that exhibits strong strand displacement. This polymerase is deficient in both proofreading (3ʹ→ 5ʹ) and nick-translation (5ʹ→ 3ʹ) nuclease activities. The protein was originally characterized and its crystal structure solved by Lorena Beese (1).

Supplied in: 
10 mM Tris-HCl, 50 mM KCl, 1.0 mM DTT, 0.1 mM EDTA, <0.1% Triton X-100 and 50% glycerol; pH 7.5 at 25°C.
 
Supplied with: 
10X PCR Buffer II (cat. no. B7140) contains 200 mM Tris-HCl, 100 mM (NH₄₎2SO₄, 100 mM KCl, 20 mM MgSO4, and 1.0% 
Triton X-100; pH 8.8 at 25°C.

Ask about low glycerol formulations.

Polymerase properties

• Storage temperature: –25°C to –15°C

Source of recombinant enzyme protein: The protein is produced by a recombinant E. coli strain carrying the Manta 1.0 DNA 
Polymerase (exo-) polymerase gene.

Unit definition: One unit is defined as the amount of polymerase required to convert 10 nmol of dNTPs into acid-insoluble 
material in 30 minutes at 65°C.

Molecular weight: 66,215 Daltons

Usage Instructions: 

Manta 1.0 DNA Polymerase exhibits strong strand displacement activity and can be used in various nucleic acid amplification 
methods, such as isothermal, whole genome, and multiple displacement amplifications.

Reaction Conditions 

Optimal reaction temperature is between 60-65°C. 

10X Reaction Buffer contains 2 mM MgSO₄ at 1X. No dNTPs are included in the buffer. 

For optimization, add MgSO₄ up to 10 mM, and Manta 1.0 Polymerase up to 1.6 U/µL.

Quality control analysis 

Unit activity was measured using a twofold serial dilution method. Dilutions of the enzyme batch were made in 1X reaction buffer 
and added to 50 µl reactions containing calf thymus DNA, 1X PCR Buffer II, 3H-dTTP and 100 µM dNTPs. The reactions were 
incubated for 10 minutes at 65°C, placed on ice and analyzed using the method of Sambrook and Russell (2). 

Protein concentration is determined by OD₂₈₀ absorbance.

Physical purity is evaluated by SDS-PAGE of concentrated and diluted enzyme solutions followed by silver-stain detection. Purity is 
assessed by comparing the aggregate mass of contaminant bands in the concentrated sample to the mass of the band corresponding 
to the protein of interest in the diluted sample.

Single-stranded exonuclease is determined in a 50 µl reaction containing a radiolabeled single-stranded DNA substrate and 10 µl
of enzyme solution incubated for 4 hours at 37°C.

Double-stranded exonuclease activity is determined in a 50 µl reaction containing a radiolabeled double-stranded DNA substrate 
and 10 µl of enzyme solution incubated for 4 hours at 37°C.

Double-stranded endonuclease activity is determined in a 50 µl reaction containing 0.5 µg of plasmid DNA and 10 µl of enzyme 
solution incubated for 4 hours at 37°C.

E. coli 16S rDNA contamination is evaluated using 5 µl replicate samples of enzyme solution that are denatured and screened in 
a TaqMan qPCR assay for the presence of contaminating E. coli genomic DNA using oligonucleotide primers corresponding to the 
16S rRNA locus.

This product is available for molecular biology applications such as: 

• Isothermal amplification 
• Whole Genome Amplification (WGA)
• Multiple displacement amplification (MDA)

References: 
1. Kiefer, et al. Structure 15 January 1997. 5, 95-108.
2. Sambrook, J. et al. (1989) Cold Spring Harbor Laboratory Press, Molecular Cloning: A Laboratory Manual., (2nd ed.), 5.40-
5.43.