- Taq DNA Polymerase
- Thermus aquaticus DNA Polymerase is a thermostable enzyme which replicates DNA at 74 degrees, remaining functional even to 95 degrees. Is used in PCR reactions and other reactions requiring synthesis of DNA at elevated temperatures. Used in PCR and DNA sequencing reactions.
- Tfi DNA Polymerase
- From Thermus flavus and is a thermostable DNA polymerase similar to Taq in its use in PCR reactions. Used in PCR and DNA sequencing reactions.
- DNA Polymerase I
- Catalyses the polymerization of deoxyribonucleotides to duplex DNA in the 5′-3′ direction. This intact polymerase possess a 3′-5′ exonuclease (editing or proofreading) function. DNA polymerase I also contains a 5′-3′ exonuclease activity enabling the enzyme to replace nucleotides in the growing strand of DNA. Used in labelling of DNA to high specific activity by nick translation, second strand cDNA synthesis, and end-labelling of DNA with protruding 3′ termini.
- DNA Polymerase I Klenow Fragment
- Proteolytic cleavage of DNA Polymerase I produces the large Klenow Fragment which retains the polymerase activity and the 3′-5′ exonuclease and strand displacement activities but lacks the 5′-3′ exonuclease activity. This enzyme is good for Filling in or labelling the 3′-ends of double stranded DNA, Second Strand cDNA synthesis, DNA sequencing by the dideoxy method. A genetically engineered Klenow Fragment which lacks the exonuclease activity all together has also been produced.
- T4 DNA Polymerase
- Catalyses the 5′-3′ synthesis of DNA from a primed single-stranded DNA template. Has a potent 3′-5′ proofreading exonuclease activity but no 5′-3′ exonuclease. Used for flush 5″ protruding ends with labelled or unlabelled dNTP’s; blunt 3′ overhangs, and in vitro mutagenesis.
- Terminal Deoxynucleotide transferase
- Purified from calf thymus and catalyses the repetitive addition of mononucleotides from dNTP to the 3’OH terminus of a DNA initiator. Single stranded DNA is preferred but not absolute. Used for tailing reactions to add complementary homopolymer tails to DNA vectors and to cDNA.
- SP6 RNA Polymerase
- Is a DNA-dependent RNA polymerase with high specificity for bacteriophage SP6 promoter sequences. Using the SP6 enzyme, only SP6 DNA or DNA cloned downstream from the SP6 promoter can serve as a template for RNA synthesis. Used for synthesis of RNA transcripts for hybridization probes, in vitro translation, RNase protection assays or RNA processing substrates.
- T7 RNA Polymerase
- Is a DNA-dependent RNA polymerase with high specificity for the T7 promoter. Applications similar to those for the SP6 Polymerase: synthesis of RNA transcripts for hybridization probes, in vitro translation, RNase protection assays or RNA processing substrates.
- AMV Reverse Transcriptase
- The AMV (Avian Myeloblastosis Virus) reverse transcriptase is a DNA polymerase which will use DNA, RNA, or RNA:DNA hybrids as a template. It requires an oligonucleotide primer and Mg or Mn. The avian enzyme possesses intrinsic ribonuclease H activity. Used for first and second strand synthesis of cDNA, primer extensions and RNA sequencing, preparation of labelled hybridization probes, and RT-PCR (reverse transcriptase-PCR amplification of mRNA to DNA).
- HIV Reverse Transcriptase
- Catalyses the synthesis of DNA in the 5′-3′ direction with a RNA template and a DNA primer, also has RNAase H activity. Can be used to produce cDNA from mRNA.
- M-MLV Reverse Transcriptase
- The Moloney Murine Leukaemia Virus – reverse transcriptase is an RNA-dependent DNA polymerase requiring a DNA primer and an RNA template to synthesize a complementary strand. M-MLV reverse transcriptase has a weaker intrinsic RNase H activity than AMV which is important for achieving full-length (long) complementary DNA. Used for first strand synthesis of cDNA, primer extensions. (A RNase minus genetically engineered enzyme is also available).
- T4 DNA Ligase
- Catalyses the joining of two strands of DNA between the 5′-phosphate and the 3′-hydroxyl groups of adjacent nucleotides either in a blunt-end or cohesive-end configuration. Will not join single stranded nucleic acids. Used for joining (ligating) double-stranded DNA molecules with cohesive or blunt ends.
- T4 Polynucleotide Kinase
- Catalyses the transfer of the gamma phosphate from ATP to the 5″ terminus of polynucleotides and to mononucleotides bearing a 3′-phosphate group. Used for 5′-labelling of single or double stranded DNA and RNA molecules for use as probes, for sequencing or for DNA-protein footprinting.
- BAL31 Nuclease
- Cleaves duplex DNA and RNA with an exonuclease activity, from both ends and produces successively shorter strands. Digestion gives mostly blunt-ends. Also acts as a single stranded exonuclease cleaving at nicks, gaps and single stranded regions of NDNA. Used for generating DNA deletions from defined endpoints (ie characterization of importance of specific DNA sequences).
- Exonuclease I
- A single-stranded DNA degrading enzyme acting processively in the 3′-5′ direction producing 5′-mononucleotides and leaving the 5′-dinucleotide intact. Used for elimination of residual single-stranded DNA containing a 3′-terminus.
- Exonuclease III
- A 3′-5′ exonuclease specific for double stranded DNA that has a blunt end, a 5′-overhang, or a nick. It will not degrade double stranded DNA with a 3′-overhang of at least four bases. The enzyme also has an endonuclease activity specific for apurinic sites, ribonuclease H activity, and a 3’phosphatase activity. Used for generating nested sets of deletions with double stranded, linear DNA and for preparation of single-stranded DNA to serve as a template for dideoxy sequencing.
- Endonuclease IV
- Multiple activities, all of which may be required for recombination or DNA repair. It hydrolyses nucleotides from both the 3′ and 5′-ends of linear double-stranded and Single -stranded DNA. Requires ATP for activity.
- Exonuclease VII
- A strict single-strand directed enzyme with 5′-3′ and 3′-5′ exonuclease activity. It is the only bi-directional exonuclease with single-strand specificity.
- Mung Bean Nuclease
- Degrades single-stranded DNA and RNA with exonuclease activity to yield 5′-phosphate terminated products. (very high concentrations will degrade double stranded DNA from both ends). Used for removal of protruding single-stranded termini in double stranded DNA and for selective cleavage of single-stranded nucleic acids in mRNA mapping experiments.
- Micrococcal Nuclease
- Catalyses cleavage of RNA and DNA to yield 3′-nucleotides (or deoxynucleotides).
- Nuclease P1
- Hydroxyzes RNA and DNA to 5′-mononucleotides. It is a random endonuclease and ultimately degrade DNA completely to deoxyribonucleotides.
- Pancreatic deoxyribonuclease
- Randomly degrades both single and double-stranded DNA producing 5′-P terminal oligonucleotides. It randomly puts nicks in double-stranded DNA in the presence of Mg and can be used for nick translation of DNA.
- Poly(A) Polymerase
- Specifically catalyses the incorporation of adenine residues onto the 3′-termini of single stranded RNA (the E. coli enzyme). The yeast enzyme will function with any oligo or polyribonucleotide.
- Ribonuclease H
- An E. coli enzyme with endonuclease activity specific for RNA hybridized to DNA. Produces 3′-OH and 5′-P terminated products. Will not degrade single-stranded nucleic acids, duplex DNA or double stranded RNA. Used for removal of the RNA strand prior to second strand cDNA synthesis and analysis of in vitro polyadenylation reaction products.
- S1 Nuclease
- Degrades single-stranded DNA and RNA with exonuclease activity yielding 5′-P terminated products. Double stranded nucleic acids of all forms are resistant to degradation except at very high enzyme concentrations. Used for removal of protruding single-stranded termini in double stranded DNA, selective cleavage of single-stranded regions of DNA and for mapping of RNA transcripts.
- Alkaline Phosphatase
- Catalyses the hydrolysis of 5′-phosphate groups from DNA, RNA, and deoxy- and ribo-nucleotides. Used for preventing re-ligation of linearised cloning vector DNA by removing the 5′-P group from both ends, removing 5′-P groups prior to end-labelling DNA with T4 Polynucleotide kinase.
- Lambda Terminase
- Recognizes the sequence of about 100 bp at the cos region of lambda DNA and cleaves the COS site to generate a 5′-protruding end with 12 bases (or 4-5 bases). Linearises the cosmid or lambda vector so clones can be mapped.