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CircLigase™ II ssDNA Ligase，单链DNA连接酶，单链DNA环化酶，1000Units
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  • 货号 CL9021K
  • 品牌 Epicentre ( 经销商 )
  • CAS号 N/A
  • 规格/包装 1000units/个
  • 单位 个
  • 储存条件 -20℃
  • 现货状态 一周
  • 数量

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说明书：CircLigase™ II ssDNA Ligase

CircLigase™ II ssDNA Ligase

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™ II ssDNA Ligase* is a thermostable enzyme 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 II 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 >15 bases, including cDNA, is circularized by CircLigase II enzyme. Under standard reaction conditions, virtually no linear concatamers or circular concatamers are produced. In addition to its activity on ssDNA, CircLigase II enzyme also has activity in ligating a single-stranded nucleic acid having a 3´-hydroxyl ribonucleotide and a 5´-phosphorylated ribonucleotide or deoxyribonucleotide.

CircLigase II has greater activity and a new Reaction Buffer for improved ligation efficiency.

Unit Definition: One unit of CircLigase II enzyme converts 1 pmol of a linear 5´-phosphorylated CircLigase II 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: 0.33 M Tris-Acetate (pH 7.5), 0.66 M potassium acetate and 5 mm DTT.

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

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


Figure 1. CircLigase™ II 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™ II 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.

*Covered by intellectual property rights licensed to Epicentre.


CircLigase™ II ssDNA Ligase
应用
·  产生单链DNA模板用于滚环复制或者滚环转录实验。
·  产生单链DNA模板用于RNA聚合酶和RNA聚合酶抑制剂测定。
CircLigase™ II ssDNA Ligase是一个热稳定性酶，催化具有5'-磷酸和3'-羟基基团的单链DNA模板的分子内连接（即环化）。与T4 DNA Ligase和Ampligase® DNA Ligase不同（连接彼此相邻的互补的DNA序列的末端）CircLigase II ssDNA Ligase可以在没有互补序列存在的情况下连接单链DNA末端。因此，该酶在将线性单链DNA连接成环状单链DNA中具有重要作用。环状的单链DNA分子可被用作滚环复制或滚环转录研究的底物。
大于15个碱基的线性单链DNA，包括cDNA，均可被CircLigase II enzyme环化。标准反应条件下，不产生线性或环状的嵌合体结构。除了对单链DNA具有活性，CircLigase II也在具有3'-羟基核糖核苷酸和5'-磷酸化的核糖核苷酸或脱氧核糖核苷酸的单链核酸的连接中具有活性的活性。
CircLigase II具有更高的活性，配备了新的改进了连接效率的缓冲液。
活力单位定义: 一个活力单位的CircLigase II酶在标准反应条件下，60°C ，1小时能够将1 pmol的线性 5´-磷酸化的标准55-mer寡核苷转换成抵抗外切酶I的环状形式。
储存缓冲液: 50%甘油含有50 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT和0.1% Triton® X-100.
10X反应缓冲液: 0.33 M Tris-乙酸盐(pH 7.5), 0.66 M醋酸钾和5 mM DTT。
质量控制: CircLigase II ssDNA Ligase不含磷酸酶、DNA外切、内切和RNA酶活性。
Product Citations
1.     Polidoros, A.N. et al. (2006) BioTechniques41, 35.
2.     Shroff , H. et al. (2005) Nano Letters5(7), 1509.
3.     Lin, C. et al. (2006) AngewandteChemie118, 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 Journal94, 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
Figure 1. CircLigase™ II ssDNA Ligase将线性单链DNA转换成环状单链DNA. A 71个碱基的单链DNA寡核苷在含有CircLigase™ II ssDNA Ligase和ATP的反应体系中被转换成环状DNA。Lane 1, DNA markers. Lane 2, 71个碱基的单链DNA. Lane 3, 环化经过腺甘酸化的中间体. Lane 4, 闭合环状的DNA通过专门切割线性DNA的外切核酸酶I进行验证。
CL9021K CircLigase™ II ssDNA Ligase 1,000 Units
CL9025K CircLigase™ II ssDNA Ligase 5,000 Units









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