Taq DNA聚合酶 (E001)

目录 规格 价格
产品描述:
本制品是94kDa的耐热性DNA聚合酶,是把Thermus aquaticus DNA Polymerase基因在大肠杆菌中进行表达后经多次纯化分离而得到 的。它与天然Taq DNA聚合酶具有相同的功能。使用本制品扩增得到的 PCR产物的3´端附有一个"A"碱基,因此可直接克隆于T载体中。配以Novoprotein独特配方的5×反应缓冲液(已添加Mg2+)可获得更加理 想的扩增结果。
产品特点:
  • 高效:以λDNA为模板,扩增长度可达8kb,可以高效扩增≤4kb片 段。
  • 灵敏:可从0.05ng人基因组DNA模板中扩增出特定基因片段。
  • 高品质:完全满足Real Time PCR的实验要求。
  • 独特配方的5×Buffer:使PCR扩增反应更加稳定、灵敏、高效。
产品用途:
  • 常规PCR鉴定;
  • 小片段目标基因克隆;
  • 平端PCR产物加A;
  • 双脱氧法测序;
  • 荧光定量PCR;
使用建议:
  • 使用本试剂扩增得到的PCR产物3´ 端有一突出"A"碱基,可直接克隆 于T载体中。
  • 如需扩增克隆较长片段建议换用Pfu DNA聚合酶。当扩增大片段 时,Taq酶的扩增效率大大低于Taq Plus酶,在>4Kb片段的PCR扩增中建议换用Taq Plus酶。
应用实例:
Taq DNA Polymerase Amplification
图例一) 50μl扩增体系中,以5ngλDNA 为模板,对500bp~6.0kb的扩增结果。
泳道M:1KB DNA Marker;泳道1:0.5kb;泳道2:1.0kb;泳道3:2.0kb;泳道4:3.0kb;泳道5:4.0kb;泳道6:6.0kb;
Taq DNA Polymerase Amplification
图例二) 50μl扩增体系中,分别以50ng~0.05ng人基因组DNA为模板,可对特定基因片段(380bp)进行很好的扩增。
泳道1:50ng;泳道2:5ng;泳道3:0.5ng;泳道4:0.05ng;泳道M:DNA Ladder 2000

产品包装:

Taq DNA 聚合酶 (5U/μl)50μl
dNTP混合液(各10mM)*200μl
5×Taq Buffer with Mg2ml
*E001-02系列不含dNTP混合液

质量保证:

经多次柱纯化,SDS-PAGE胶检测仅可见清晰单一的目的条带;PCR方法检测无大肠杆菌DNA残留,无核酸内、外切酶污染。

活性定义:

在74℃条件下,30分钟内催化10nmol dNTP的掺入反应成为酸不溶性物质所需的酶量为一个单位。

常用反应体系(50μl)
5×Taq Buffer10μl
上游引物0.2-1.0μM(终浓度)
下游引物0.2-1.0μM(终浓度)
dNTP(各10mM)1.0μl
模板1-50ng(质粒)
10ng-1μg(基因组)
Taq0.25μl(1.25U)
ddH2O至50μl

常用PCR循环:

当扩增片段<3K:
94 C 1分钟30秒
30次循环94℃,20秒/57℃
20秒/72℃
根据产物长度调整, 60秒/kb
72℃5分钟
4℃保温
当扩增片段≥3K(推荐引物长度≥30bp):
94 C20秒
30次循环 94℃,5秒/68℃
根据产物长度调整, 60秒/kb
72℃5分钟
4℃保温
参考文献

  • Wu J, Gao B, Zhu S.
    Single-point mutation–mediated local amphipathic adjustment dramatically enhances antibacterial activity of a fungal defensin.
    The FASEB Journal(2016)

  • Wang X, Umetsu Y, Gao B, et al.
    Mesomartoxin, a new K v 1.2-selective scorpion toxin interacting with the channel selectivity filter.
    Biochemical pharmacology, 2015, 93(2): 232-239.(2015)

  • Shunyi Zhu, Lan Luo, Ping Li, Bin Gao, Limei Zhu, Yuzhe Yuan .
    DrTx(1-42), a C-terminally truncated analogue of drosotoxin, is a candidate of analgesic drugs.
    Biochemical Pharmacology (2011)

  • Shunyi Zhu , Bin Gao , Meichun Deng , Yuzhe Yuan , Lan Luo, Steve Peigneur ,Yucheng Xiao, Songping Liang , Jan Tytgat .
    Drosotoxin, a selective inhibitor of tetrodotoxin-resistant sodium channels.
    Biochemical Pharmacology(2010)

  • Zhaohua Hou,Qiuju Han,Cai Zhang,Zhigang Tian,Jian Zhang.
    miR146a impairs the IFN‐induced anti‐HBV immune response by down‐regulating STAT1 in hepatocytes.
    Liver International(2013)

  • Ping Li, Shunyi Zhu.
    Mutational analysis of the analgesic peptide DrTx(1-42) revealing a functional role of the amino-terminal turn.
    PLoS ONE(2012)

  • Zhenting Zhang, Shunyi Zhu.
    Functional role of charged residues in drosomycin, a Drosophila antifungal peptide..
    Developmental and Comparative Immunology 34 (2010) 953–958 (2010)

  • Bin Gao, Shunyi Zhu.
    Identification and characterization of the parasitic wasp Nasonia defensins: positive selection targeting the functional region.
    Developmental and Comparative Immunology 34 (2010) 659–668(2010)

  • Hu D, Hu S, Wan W, et al.
    Effective Optimization of Antibody Affinity by Phage Display Integrated with High-Throughput DNA Synthesis and Sequencing Technologies.
    PloS one, 2015, 10(6): e0129125(2015)

  • Bin Gao, Shunyi Zhu.
    Characterization of a hymenoptaecin-like antimicrobial peptide in the parasitic wasp Nasonia vitripennis.
    Process Biochemistry (2010)

  • Caihuan Tian ,Lei Wang, Gongyin Ye ,Shunyi Zhu.
    Inhibition of melanization by a Nasonia defensin-like peptide: implications for host immune suppression.
    Journal of Insect Physiology (2010)

  • Yuzhe Yuan, Lan Luo , Steve Peigneur, Jan Tytgat , Shunyi Zhu.
    Two recombinant depressant scorpion neurotoxins differentially affecting mammalian sodium channels..
    Toxicon (2010)

  • Caihuan Tian, Bin Gao, Maria del Carmen Rodriguezb, Humberto Lanz-Mendoza,Bo Ma, Shunyi Zhu.
    Gene expression, antiparasitic activity, and functional evolution of the drosomycin family..
    Molecular Immunology(2008)

  • Wang X, Gao B, Zhu S.
    A single-point mutation enhances dual functionality of a scorpion toxin.
    Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology(2016)

  • Kuanyu Zhu, Bin Gao, Shunyi Zhu .
    Characterization of a chimeric antimicrobial peptide uncovers evolutionary significance of exon-shuffling.
    Biochemical and Biophysical Research Communications 428(2013)360-364 (2013)

  • Yong-Gang Xie, Chao Luan, Hai-Wen Zhang, Fei-Fei Han, Jie Feng, Young-Jun Choi, Denis Groleau, Yi-Zhen Wang .
    Effects of Thioredoxin: SUMO and Intein on Soluble Fusion Expression of an Antimicrobial Peptide OG2 in Escherichia coli.
    Protein & Peptide Letters(2013)

  • Ping Li, Shunyi Zhu.
    Molecular design of new sodium channel blockers.
    Biochemical and Biophysical Research Communications 414(2011)321-325 (2011)

  • Yi Cao, Li Liao,Xue-wei Xu,Aharon Oren,Min Wu.
    Aldehyde dehydrogenase of the haloalkaliphilic archaeon Natronomonas pharaonis and its function in ethanol metabolism.
    Extremophiles(2008)

  • Yuzhe Yuan, Bin Gao, Shunyi Zhu.
    Functional expression of a Drosophila antifungal peptide in Escherichia coli..
    Protein Expression and Purification 52 (2007) 457–462(2007)

  • Chen X, Shi J, Chen R, et al..
    Molecular chaperones (TrxA, SUMO, Intein and GST) Mediating Expression, Purification and Antimicrobial Activity Assays of Plectasin in Escherichia coli[J]..
    Biotechnology and applied biochemistry(2014)

  • Xiaohui LIU1, Xiaodong PENG2, Zhenzhen HU 2, Qingmei ZHAO 2, Jian HE2, Junhe LI2, Xiaojun ZHONG2.
    Effects of over-expression of ANXA10 gene on proliferation and apoptosis of hepatocellular carcinoma cell line HepG2..
    J Huazhong Univ Sci Technol(2012)

  • 李见, 王露, 李玉云..
    重组表达人IDO 基因慢病毒载体的构建及鉴定[J]..
    蚌埠医学院学报(2014)

  • 李摇贞, 徐摇庆, 张摇慧.
    慢病毒介导的TGFβ-R2 shRNA 抑制人 LECs 细胞TGFβ-R2的表达.
    (2014)

  • 曲利娟, 苏娟娟, 梁莉.
    DR-nm23 基因慢病毒表达载体的构建及其在人结直肠癌 SW620 细胞中的表达.
    世界华人消化杂志(2011)

  • 韩世伟,张阳德,万小平,陈玉祥.
    重组外源性人热休克蛋白 70 基因真核表达载体的构建及鉴定 .
    中国组织工程研究(2010)

  • 汪月霞, 王 忠, 刘全军, 赵会杰, 顾蕴洁, 钱晓旦, 袁志良.
    cpt1基因与水稻根负向光性运动的关系.
    作物学报(2009)
For research use ONLY.