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Enzymes for Molecular Biology

Enzymes for Molecular Biology: Enzymes for Molecular Biology

CircLigase™ ssDNA Ligase

Production of ssDNA templates for rolling-circle replication or rolling-circle transcription experiments. Also used in ssDNA templates for RNA polymerase and RNA polymerase inhibitor assays

BioSearch Tech (Lucigen/Epicentre)

Catalogue No.DescriptionPack SizePriceQty
CL4111K CircLigase™ ssDNA Ligase 1000 units £159.00 Quantity Add to Order
CL4115K CircLigase™ ssDNA Ligase 5000 units £660.00 Quantity Add to Order

CircLigase™ ssDNA Ligase

Production of ssDNA templates for rolling-circle replication or rolling-circle transcription experiments. Also used in ssDNA templates for RNA polymerase and RNA polymerase inhibitor assays

BioSearch Tech (Lucigen/Epicentre)

Applications

  • Production of ssDNA templates for rolling-circle replication or rolling-circle transcription experiments.
  • Production of ssDNA templates for RNA polymerase and RNA polymerase inhibitor assays.

CircLigase™ ssDNA Ligase* is a thermostable ATP-dependent ligase that catalyzes intramolecular ligation (i.e. circularization) of ssDNA templates having a 5´-phosphate and a 3´-hydroxyl group. In contrast to T4 DNA Ligase and Ampligase® DNA Ligase, which ligate DNA ends that are annealed adjacent to each other on a complementary DNA sequence, CircLigase ssDNA Ligase ligates ends of ssDNA in the absence of a complementary sequence. The enzyme is therefore useful for making circular ssDNA molecules from linear ssDNA. Circular ssDNA molecules can be used as substrates for rolling-circle replication or rolling-circle transcription.

Linear ssDNA of >30 bases is circularized by CircLigase enzyme. Under standard reaction conditions, virtually no linear concatamers or circular concatamers are produced. In addition to its activity on ssDNA, CircLigase enzyme also has activity in ligating a single-stranded nucleic acid having a 3´-hydroxyl ribonucleotide and a 5´-phosphorylated ribonucleotide or deoxyribonucleotide.

 

 

Figure 1. CircLigase™ ssDNA Ligase converts linear ssDNA to circular ssDNA. A 71-base ssDNA oligo was converted to a circular DNA form in a reaction containing CircLigase™ ssDNA Ligase and ATP. Lane 1, DNA markers. Lane 2, 71-base ssDNA. Lane 3, circularization proceeds through an adenylated intermediate. Lane 4, the closed circular nature of the reaction product was confirmed by treating the reaction with exonuclease I, which specifically digests linear DNA.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

CircLigase™ ssDNA Ligase

Production of ssDNA templates for rolling-circle replication or rolling-circle transcription experiments. Also used in ssDNA templates for RNA polymerase and RNA polymerase inhibitor assays

BioSearch Tech (Lucigen/Epicentre)

protocol can be be downloaded from the manufacturers website here

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

CircLigase™ ssDNA Ligase

Production of ssDNA templates for rolling-circle replication or rolling-circle transcription experiments. Also used in ssDNA templates for RNA polymerase and RNA polymerase inhibitor assays

BioSearch Tech (Lucigen/Epicentre)

Product Citations

  1. Polidoros, A.N. et al. (2006) BioTechniques 41, 35.
  2. Shroff , H. et al. (2005) Nano Letters 5(7), 1509.
  3. Lin, C. et al. (2006) Angewandte Chemie 118, 7699.
  4. Korlach, J. et al. (2008) Proc. Natl. Acad. Sci. USA 105, 1176.
  5. McArthur, M. and Bibb, M.J. (2008) Proc. Natl. Acad. Sci. USA 105, 1020.
  6. Shroff , H. et al. (2008) Biophysical Journal 94, 2179.
  7. Kuhn, H. and Frank-Kamenetskii, M.D. (2008) Nucleic Acids Res. 36, e40.
  8. Murakami, T. et. al. (2008) Nucleic Acids Res. Doi:10.1093/nar/gkn1014

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

CircLigase™ ssDNA Ligase

Production of ssDNA templates for rolling-circle replication or rolling-circle transcription experiments. Also used in ssDNA templates for RNA polymerase and RNA polymerase inhibitor assays

BioSearch Tech (Lucigen/Epicentre)

Unit Definition: One unit of CircLigase enzyme converts 1 pmol of a linear 5´-phosphorylated CircLigase Standard 55-mer Oligo into an exonuclease I-resistant circular form in 1 hour at 60°C under standard assay conditions.

Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton® X-100.

10X Reaction Buffer: 0.5 M MOPS (pH 7.5), 0.1 M KCl, 50 mM MgCl2, and 10 mM DTT. The Reaction Buffer does not contain
ATP or MnCl2 which must be added to the reaction at final concentrations of 0.05 mM ATP and 2.5 mM MnCl2. A 2.5-mM solution of ATP and a 50-mM solution of MnCl2 are included.

Quality Control: CircLigase ssDNA Ligase is free of detectable phosphatase, DNA exo- and endonuclease, and RNase activities

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200