DNA Modifying Enzymes and Cloning Technologies

Blunt/TA Ligase Master Mix is a ready-to-use solution of T4 DNA Ligase, proprietary ligation enhancer, and optimized reaction buffer. This master mix is specifically formulated to improve ligation and transformation of both blunt-end and single-base overhang substrates.

• Simplifies reaction set-up, ensures an optimized ratio of enzyme and buffer components
• No thawing is necessary as it remains liquid during storage at -20°C
• Ligations for subcloning can be carried out in small volumes with low DNA concentrations, allowing users to conserve precious DNA samples and directly transform many strains of chemically competent E. coli without dilution
• Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart

• Ideal for in vitro digestion of dsDNA
• Compatible with EnGen^® sgRNA Synthesis Kit, S. pyogenes (NEB #E3322S) and the EnGen Mutation Detection Kit (NEB #E3321S)
• For help with oligo design, try EnGen sgRNA Template Oligo Designer – now included in the NEBioCalculator^® Tool

• Endonuclease that nonspecifically cleaves DNA to release di-, tri- and oligonucleotide products with 5′-phosphorylated and 3′-hydroxylated ends
• Degrades DNA template in transcription reactions
• Removes contaminating genomic DNA from RNA samples
• Isolated from a recombinant source

Characteristics of Hi-T4 DNA Ligase:

• Hi-T4 DNA Ligase is an engineered variant of T4 DNA Ligase with improved thermostability compared to T4 DNA Ligase
• Capable of joining blunt end and cohesive end termini, as well as repairing single stranded nicks in duplex DNA and some DNA/RNA hybrids
• Supplied with both T4 DNA Ligase Buffer and StickTogether^™ DNA Ligase Buffer
• Other T4 DNA Ligase products include Quick Ligation Kit, Salt-T4, Instant Sticky-end Ligase Master Mix and Blunt/TA Ligase Master Mix
• Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart or DNA Ligase Selection Chart

Specifically formulated to rapidly ligate cohesive-end (2–4 bp) substrates and improve transformation, Instant Sticky-end Ligase Master Mix is a ready-to-use 2X solution of T4 DNA Ligase and a proprietary ligation enhancer in an optimized reaction buffer.

• No incubation time necessary to achieve ligation
• Efficiencies sufficient for successful sub-cloning of sticky-end substrates
• No thawing of the master mix is required as liquid state is maintained when stored at -20°C
• Simplified reaction setup with optimized ratio of enzyme and buffer components
• Ligations for subcloning can be carried out in small volumes with low DNA concentrations, allowing users to conserve precious DNA samples and directly transform many strains of chemically competent E. coli without dilution
• Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart

Quick CIP from NEB is a heat-labile version of calf intestinal alkaline phosphatase (CIP) purified from a recombinant source.

Features of Quick CIP, NEB:

• Rapid and irreversible heat inactivation eliminates the unwanted activity
• Improved storage stability versus native enzyme
• Faster reaction setup (no supplemental additives like zinc required) and shorter incubation time
• Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required)
• Less enzyme required (high specific activity), resulting in a lower cost per reaction
• No need for multiple phosphatases (Quick CIP removes 5′- and 3′- phosphates from DNA, RNA and dNTPs )
• Active on unincorporated dNTPs in PCR products – improve DNA sequencing and SNP analysis
• Recombinant for purity, consistency and value

The Quick Ligation™ Kit enables ligation of cohesive end or blunt end DNA fragments in 5 minutes at room temperature. (25°C )

• Fast – 5 minutes for cohesive or blunt ends
• Convenient – ligation performed at room temperature
• Flexible – suitable for all common ligation reactions
• Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart

Characteristics of Salt-T4 DNA Ligase:

• Salt-T4 DNA Ligase is an engineered variant of T4 DNA Ligase with improved salt tolerance
• Capable of joining blunt end and cohesive end termini, as well as repairing single stranded nicks in duplex DNA and some DNA/RNA hybrids.
• Supplied with both T4 DNA Ligase Buffer and StickTogether™ DNA Ligase Buffer.
• Other T4 DNA Ligase products include Quick Ligation Kit, Hi-T4, Instant Sticky-end Ligase Master Mix and Blunt/TA Ligase Master Mix
• Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart or DNA Ligase Selection Chart

Shrimp Alkaline Phosphatase (rSAP) is a heat labile alkaline phosphatase purified from a recombinant source.

• Rapid and irreversible heat inactivation eliminates unwanted activity
• Improved storage stability versus native enzyme
• Faster reaction setup (no supplemental additives like zinc required)
• Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required)
• Less enzyme required (high specific activity), resulting in a lower cost per reaction
• No need for multiple phosphatases (rSAP removes 5´- and 3´- phosphates from DNA, RNA and dNTPs )
• Active on unincorporated dNTPs in PCR products – improves DNA sequencing and SNP analysis
• Recombinant for purity, consistency and value

Characteristics of T4 DNA Ligase, NEB:

• Catalyzes the formation of a phosphodiester bond between juxtaposed 5′ phosphate and 3′ hydroxyl termini in duplex DNA or RNA.
• This enzyme will join blunt end and cohesive end termini as well as repair single-stranded nicks in duplex DNA and some DNA/RNA hybrids (1).
• T4 DNA ligase will seal nicks for these DNA substrates.
• The industry standard for performance and quality
• Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart