Product Number: LG02

Shipping and Storage

Store at -30～-15 ℃ and transport at ≤ 0℃.

Components

Description

T4 DNA Ligase catalyzes the formation of phosphodiester bonds between adjacent 5′- phosphate and 3′ – hydroxyl ends on double stranded DNA or RNA. This enzyme can not only catalyze connections between flat or sticky ends, but also repair single strand cleavage in double stranded DNA, RNA, or DNA/RNA hybrid double strands.

Application

1. The connection between DNA fragments and carrier DNA.
2. The connection between DNA fragments and Linker or Adaptor DNA.

Unit definition

In a 20μl linkage reaction system, when 6μg of λDNA-Hind III decomposition product was reacted at 16℃ for 30 minutes, more than 50% of DNA fragments were linked, and the required enzyme quantity was defined as one active unit (U).

Product Number: LG0N

Shipping and Storage

Store at -20℃ and transport on dry ice.

Components

Description

T4 DNA Ligase is isolated and purified from Escherichia coli expressing the T4 DNA Ligase gene after induction and expression. It catalyzes the binding reaction of adjacent DNA strands with 5’phosphate groups and 3’hydroxyl groups via phosphodiester bonds. This enzyme can catalyze the connection of flat or sticky end DNA, repairing single stranded incisions in double stranded DNA, RNA, and DNA/RNA hybridization, but has no activity for single stranded nucleotides.

Application

Mainly used for adapter connection during the construction process of NGS Chinese library.

Unit definition

1U refers to the amount of enzyme required to convert 1nmol [32PPi] into Norit absorbable form within 20 minutes at 37°C in the ATP-PPi exchange reaction, equivalent to approximately 200 viscous terminal junction units.

Product Number: T4K01

Shipping and Storage

-20°C.

Components

Description

T4 PNK is a polynucleotide 5 ‘hydroxy kinase that catalyzes ATP γ- Transfer and exchange of phosphonic acid to double stranded/single stranded DNA or RNA, as well as single nucleotides with 3 ‘phosphate groups at 5’ – hydroxyl groups: 5 ‘- OH+NTP5’ ↔ P+NDP; This enzyme also has 3 ‘- phosphatase activity, which hydrolyzes the 3’ – phosphate group from the 3 ‘- phosphate end of the oligonucleotide, deoxygenated 3’ – monophosphate nucleoside, and deoxygenated 3 ‘- diphosphate nucleoside.

Application

1. Phosphorylation of the 5 ‘end of primers or PCR products for linkage reactions.
2. Phosphorylation of the 5 ‘end of the synthesized DNA connector (Linker) for connection reaction.
3. Labeling of the 5 ‘end of DNA and RNA, used as an oligonucleotide probe.

Unit Definition

The amount of enzyme required to mix 1 nmol of [γ-32P]ATP with acid insoluble precipitate is defined as 1 active unit, using Micrococcal Nuclease treated calf thymus DNA as substrate, at 37 ℃, pH 7.6, within 30 minutes.

Quality control

After multiple column purification, only a clear and single target band was visible in SDS-PAGE gel detection. The PCR method detected no residual Escherichia coli DNA, and no contamination of nucleic acid endonucleases, exonucleases, phosphatases, and RNA enzyme activities.

Product Number: PC71

Shipping and Storage

-20°C

Components

Description

T4 Polynucleotide Kinase, abbreviated as T4 PNK, is expressed in Escherichia coli. The expressed gene is derived from T4 bacteriophage and can catalyze the phosphorylation of ATP γ- The 5′- hydroxy terminus and 3′ – monophosphate nucleosides of the nucleotide chain (double stranded or single stranded DNA or RNA) undergo transfer and exchange, while also possessing 3’phosphatase activity, which can hydrolyze the 3′-phosphate group from the 3′ phosphate terminus of the oligonucleotide, deoxygenated 3′-monophosphate nucleoside, and deoxygenated 3′- diphosphate nucleoside.The T4 polynucleotide kinase can be used for 5′ end labeling or phosphorylation of oligonucleotides, DNA, or RNA; Catalyze the phosphorylation of single nucleotide 5’and remove the 3′ terminal phosphate group. Heating this product at 75℃ for 10 minutes can inactivate it, and adding EDTA can also inactivate it. Metal ion chelating agents, phosphates, ammonium ions, KCl greater than 50 mM, and NaCl can significantly inhibit its activity.

Unit definition

The amount of enzyme required to transfer the 1 nmolγ-phosphate group on ATP to the 5′- OH terminal of DNA within 30 minutes at 37℃ is defined as 1 active unit.

Quality control

After multiple column purification, the purity was detected by SDS-PAGE to be over 99%; After testing, there was no contamination of nucleic acid endonuclease, exonuclease, phosphatase, and RNA enzyme activities.

Description

T4 Gene 32 Protein is a single-stranded DNA (ssDNA) binding protein required for bacteriophage T4 replication and repair. It cooperatively binds to and stablizes transiently formed regions of ssDNA and plays an important structural role during T4 phage replication. It also has been used extensively to stabilize and mark regions of ssDNA for electron microscopic examination of intracellular DNA structures. Recently, it has been shown to improve restriction enzyme digestion, improve the yield and efficiency of reverse transcription reactions during RT-PCR, enhance T4 DNA polymerase activity, as well as increase the yield of PCR products.

Applications

RT-PCR & cDNA Synthesis

Features

• Stabilization and marking of ssDNA structures
• Increase yield and processivity of RT during RT-PCR
• Increase yield and specificity of PCR products from soil samples

Transportation

on blue ice

Storage

at -20°C for 24 months

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