DNA Pol I Large fragment (Klenow) was originally derived as a proteolytic product of E. coli DNA polymerase. It retains polymerase activity, but lacks both 5’ —> 3’ and 3' —> 5’ exonuclease activity. Generates probes using random primers Dideoxy sequencing Random priming labeling Moderate strand displacement activity Not suitable for generating blunt ends Product Notes Klenow Fragment (3´→ 5´ exo-) is not suitable for generating blunt ends because it lacks the 3´→ 5´ exonuclease necessary to remove non-templated 3´ additions. When Klenow Fragment (3´→ 5´ exo-) is used to sequence DNA using the dideoxy method of Sanger et al., 1 unit/5 µl reaction volume is recommended.

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Description Phi29 DNA polymerase is a DNA polymerase cloned from Bacillus subtilis phage phi29 and expressed by E. coli using gene recombination technology. This product has the ability of efficient continuous DNA synthesis and chain replacement, and has the function of 3 '→ 5' exonuclease reading, with high fidelity. This product can be applied to replication reactions requiring strong replacement and continuous synthesis and high fidelity replication under medium temperature conditions, such as plasmid replication, whole genome amplification, etc. Unit Definition At 30°C, within 10 minutes, the amount of enzyme required to add 0.5 pmol of deoxynucleotides to the acid insoluble precipitate is defined as 1 active unit (U). Thermal inactivation Incubation at 65°C for 10 minutes can inactivate. Quality Control After multiple column purification, the purity of SDS-PAGE was more than 95%; No endonuclease activity and no host residual DNA were detected. The enzyme buffer contains reductant DTT to ensure its maximum enzyme activity. If the buffer is not fresh or after repeated freezing and thawing, 4 mm DTT should be added before use. Storage Condition -20°C Components Phi29 DNA Polymerase, 10 U/μl 10x Phi29 Reaction Buffer BSA, 10 mg/ml

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RNase H, Recombinant is a ribonucleic acid endonuclease that specifically breaks down RNA strands in RNA DNA hybrids, and cannot digest single or double stranded DNA.This product can be used for the detection of DNA-RNA hybrids, removing mRNA during the synthesis of the second strand of cDNA, and removing the poly (A) tail of mRNA hybridized to poly (dT). Features This product is a thermally stable RNase H enzyme that still exhibits RNase H activity after incubation at 70°C or above for 10 minutes. Unit definition Using poly (rA)·poly (dT) as the substrate, the enzyme required to produce 1nmol of acid soluble substance within 20 minutes at 37°C is defined as 1 active unit (U) Storage Buffer Tris-HCl (pH7.5) 25mM，NaCl 30mM，DTT 1mM，EDTA 0.5mM，Glycerol 50%. Shipping and Storage -20°C. Components SKU RNH01-01 RNase H, Recombinant, 60U/μl 10μl 5×RNase H Reaction Buffer 300μl

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Description This product is expressed by Escherichia coli, and the source of the expressed gene is T4 bacteriophage. Due to the simultaneous activity of 5'→3' DNA polymerase and 3'→5' DNA exonuclease, T4 DNA polymerase can be used to flatten the 5' protruding end or flatten the 3' protruding end. It can also be used for labeling DNA probe synthesis through displacement reactions, analyzing the starting point of mRNA transcription through primer elongation, synthesizing the second strand during site-specific mutagenesis, and cloning PCR products that do not rely on linkage reactions.The 3'→5' DNA exonuclease activity of this T4 DNA polymerase is about 100-1000 times higher than Klenow Fragment, and it has higher activity for single stranded DNA than double stranded DNA. This enzyme does not contain exonuclease activity of 5'→3' DNA, and can be inactivated by heating at 70°C for 10 minutes. Metal ion chelating agents can inhibit its activity. Unit definition The amount of enzyme required to incorporate 10 nmol of whole nucleotides into acid insoluble precipitate is defined as 1 active unit (U) within 30 minutes, using thermally denatured calf thymus DNA as a template/primer, at 37°C and pH 8.8. Quality control 2U of this enzyme reacted with 1μg of Closed Circular (RFI) pBR322 DNA at 37℃ for 16 hours, and the electrophoresis bands of the DNA remained unchanged. Shipping and Storage -20°C Components SKU TDP01-01 TDP01-02 T4 DNA Polymerase (3U/μl) 50 μl 250 μl 10×T4 DNA Polymerase Reaction Buffer 1 ml 4x1 ml

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T4 Polynucleotide Kinase, abbreviated as T4 PNK, is expressed in Escherichia coli. The 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). T4 PNK also possesses 3'-phosphatase activity, which hydrolyze the 3'-phosphate group from the 3'-phosphate terminus of the oligonucleotide, deoxygenated 3'-monophosphate nucleoside, and deoxygenated 3'-diphosphate nucleoside. The T4 PNK can be used for 5' end labeling or phosphorylation of oligonucleotides, DNA, or RNA; the phosphorylation of 5' single nucleotide or removal of the 3' terminal phosphate group. Heating this product at 75°C for 10 minutes or adding EDTA can inactivate it. Ion chelating agents, phosphates, ammonium ions, KCl ≥50 mM, and NaCl can significantly inhibit its activity. Unit Definition At 37°C, within 30 minutes, the amount of enzyme required for the transfer of 1 nmol of γ- Phosphate group on ATP to the end of 5′-OH on DNA is defined as 1 active unit. Quality Control After multiple column purification, the purity of SDS-PAGE was more than 99%; No contamination of endonuclease, exonuclease, phosphatase and RNase activities was detected. Precautions Because the ammonium salt can strongly inhibit the activity of T4 polynucleotide kinase, the DNA obtained from ammonium salt precipitation cannot be used for the labeling reaction of T4 polynucleotide kinase. PEG can promote the rate and efficiency of phosphorylation reaction, and PEG should be added to the exchange reaction system. When using, the enzyme should be stored in an ice box or on an ice bath, and should be stored at -20°C immediately after use.

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Description HiFi PCR Mix for NGS is a premixed system consisting of hot-start enzyme, PCR Buffer, dNTPs, Mg2+, and PCR stabilizers and enhancers. It has high fidelity, high extensibility, and low preference. For complex DNA templates (such as high GC content template), it has a balanced amplification efficiency. This product is particularly suitable for the amplification of multiplex PCR during the construction of NGS library. The highly efficient hot-start enzyme contained in this product does not have polymerase activity at room temperature, thus effectively avoiding non-specific amplification. The combination of a unique buffer system and a hot-start enzyme significantly improves the efficiency of the PCR, resulting in a wider range of amplification. This product effectively increases the amplification efficiency of high GC or high AT regions in the genome, reduces the amplification preference and increases the coverage area of sequencing. Precautions This product cannot use with primers with modifications. Invert gently to mix well before use. Avoid foaming. Use after brief centrifuge. Avoid freezing and thawing repeatedly. Repeated freezing and thawing will comprise the performance of this product. Store at 2-8°C if used frequently.

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