Cloning Enzymes

In molecular cloning, recombined DNA is generated and inserted into a host for replication. This process requires reliable and efficient enzymes as well as high quality reagents. GoldBio offers multiple cloning enzymes including our Hot Start Pfu DNA polymerases, T4 DNA ligases, T4 DNA polymerases, T4 Polynucleotide Kinase (PNK) to help you achieve cloning success. Here you will also find detailed and comprehensive protocols for different stages of the cloning process.

Showing 1-6 of 6 items.

Pfu 2x DNA Polymerase Master Mix

There is no image for Pfu 2x DNA Polymerase Master Mix Pfu 2x DNA Polymerase Master Mix
Catalog ID Size Pricing
P-660-200 200 reactions $ 247.00

Description

Pfu DNA polymerase is a heat stable DNA polymerase which has 5´→ 3´ DNA polymerase and 3´→ 5´ exonuclease (proofreading) activities. Pfu DNA polymerase retains the high fidelity, sensitivity and processivity with an error rate six-fold lower than Taq DNA polymerase, and significantly lower than the error rates of most other proofreading enzymes or DNA polymerase mixtures. This product is supplied with an optional 5X GC Enhancer that increases the amplification of GC rich templates up to 84%.

Product Specifications

Catalog ID P-660
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P-660-200 200 reactions $ 247.00

T4 DNA Ligase - 2000 units/µL

There is no image for T4 DNA Ligase - 2000 units/µL T4 DNA Ligase - 2000 units/µL
Catalog ID Size Pricing
T-411-100 100 KU (2000 units/µl) $ 259.00
T-411-400 400 KU (2000 Units/μl) $ 335.00

Description

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5’-phosphate and 3’-hydroxyl termini in duplex DNA or RNA. This enzymes joins DNA fragments with either cohesive or blunt termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.

Product Specifications

Catalog ID T-411
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T-411-100 100 KU (2000 units/µl) $ 259.00
T-411-400 400 KU (2000 Units/μl) $ 335.00

T4 DNA Ligase - 400 units/µL

There is no image for T4 DNA Ligase - 400 units/µL T4 DNA Ligase - 400 units/µL
Catalog ID Size Pricing
T-410-100 100 KU (400 Units/μl) $ 259.00

Description

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5’-phosphate and 3’-hydroxyl termini in duplex DNA or RNA. This enzymes joins DNA fragments with either cohesive or blunt termini as well as repair single stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.

Product Includes

  • T4 DNA Ligase
  • 10x T4 DNA Ligase Reaction Buffer

Purity: ≥99% (assessed by SDS-PAGE with Coomassie blue staining)

Concentration: 400 unit/μl
One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA (250 ng/µl) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X T4 DNA ligase reaction buffer.

Source: E. coli strain expressing a recombinant clone

Inhibition and Inactivation:
Inhibitors: metal chelators, phosphate and ammonium ions, KCl and NaCl at a concentration higher than 50 mM.
Inactivated by heating at 70°C for 15 min or by addition of EDTA.

Quality Control
Endonuclease Activity (Nicking): 1 µg of supercoiled plasmid DNA is incubated with 20 units of T4 DNA Ligase in 1x Ligase buffer for 2 hours at 37ºC. Following incubation, the supercoiled DNA is visualized on an ethidium bromide stained agarose gel. No visible nicking or cutting of DNA was found.

Product Specifications

Catalog ID T-410
Storage/Handling Store at -20°C.
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T-410-100 100 KU (400 Units/μl) $ 259.00

T4 DNA Polymerase

There is no image for T4 DNA Polymerase T4 DNA Polymerase
Catalog ID Size Pricing
T-412-500 500 Units $ 171.00

Description

No description available

Product Specifications

Catalog ID T-412
Storage/Handling Store desiccated at -20°C.
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T-412-500 500 Units $ 171.00

T4 Polynucleotide Kinase (PNK)

There is no image for T4 Polynucleotide Kinase (PNK) T4 Polynucleotide Kinase (PNK)
Catalog ID Size Pricing
T-413-2500 2500 Units $ 178.00

Description

T4 Polynucleotide Kinase (PNK) catalyzes the transfer of the γ-phosphate from ATP to the 5’-OH group of single- and double-stranded DNAs and RNAs, oligonucleotides or nucleoside 3’-monophosphates. In the presence of ADP, T4 PNK exhibits 5’-phosphatase activity and catalyzes the exchange of phosphate group between 5’-P-oligonucleotides/polynucleotides and ATP. The enzyme is also a 3’-phosphatase.

NOTE: T4 PNK requires ATP for activity, but it is not supplied with ATP because it interferes with radiolabeling reactions.

Product Includes

  • T4 Polynucleotide Kinase (PNK)
  • 10x T4 PNK Reaction Buffer

Product Specifications

Catalog ID T-413
Storage/Handling Store at -20°C.
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T-413-2500 2500 Units $ 178.00

Taq DNA Ligase

There is no image for Taq DNA Ligase Taq DNA Ligase
Catalog ID Size Pricing
T-960-10 10,000 units $ 218.00

Description

Taq DNA Ligase catalyzes the formation of a phosphodiester bond in duplex DNA containing adjacent 5′-phosphoryl and 3′-hydroxyl termini, using NAD+ as a cofactor. The ligation will occur only if the oligonucleotides are perfectly paired to the complementary target DNA and have no gaps between them; therefore, a single-base substitution can be detected. Taq DNA Ligase is active at elevated temperatures (45-70°C).

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T-960-10 10,000 units $ 218.00

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