Invitrogen Platinum II Taq热启动 DNA 聚合酶

使用通用退火温度,简化PCR流程

Invitrogen Platinum II Taq 热启动DNA聚合酶旨在让您更快地达到研究目的。使用通用退火温度,可减少反应优化步骤,并可实现对不同 PCR 反应同时进行扩增。将创新型缓冲液、高性能 Taq DNA 聚合酶,以及出色热启动技术独创性地结合在一起,可获得出色的 PCR 结果,即使在最严苛的实验应用中。

产品优势

  • 通用引物退火温度 (60°C)——减少了繁琐的PCR优化步骤,并可实现对不同 PCR 反应同时进行扩增
  • 快速DNA合成速度和抑制剂耐受性——使用经改造Taq 聚合酶
  • Platinum 热启动技术——可提供优异的特异性、灵敏度及产量;并可实现在室温条件下配制反应体系
  • 绿色缓冲液——可实现 PCR 产物直接凝胶上样电泳,有助于减少移液错误

 

专为更好的PCR扩增而设计

点击以下按钮,了解Platinum II Taq 热启动DNA聚合酶的优点。

interactive pcr

关于该酶特性的视频

Platinum II Taq 热启动 DNA 聚合酶的优势

使用常规PCR试剂进行PCR扩增时,由于不同引物的退火温度和延伸时间不一致,需要针对每个DNA片段的扩增设计特定的PCR扩增方案。因此,不同的PCR反应不能同时在一个PCR运行中进行扩增。使用 Platinum II Taq 热启动 DNA 聚合酶,可以使用通用退火温度,且延伸时间可按照最长片段计算,因而可实现不同的 PCR 反应同时进行扩增。此外,Platinum II Taq 热启动 DNA 聚合酶是一种“快速”DNA 聚合酶;因此,这种新一代 DNA 聚合酶结合通用 PCR 扩增方案后,可在 30 分钟内通过快速扩增循环完成所有 PCR 反应。

Platinum2TaqHot-StartDNA-Polymerase-cycle-together

图1:可同时进行PCR扩增,极大的节省时间。使用常规PCR试剂进行PCR扩增时,由于不同引物的退火温度和延伸时间不一致,所以需要针对每个DNA片段设计特定的扩增方案。因此,使用传统的PCR试剂,多个目标片段通常不能在同一个PCR运行中一起扩增。使用 Platinum II Taq 热启动 DNA 聚合酶,可以使用通用退火温度,且延伸时间可按照最长片段计算,因而可实现不同的 PCR 反应同时进行扩增。此外,Platinum II Taq 热启动 DNA 聚合酶是一种快速的 DNA 聚合酶,只需 30 分钟便可获得 PCR 结果。

Time saving enabled by assay co-cycling

图2:Platinum II Taq 热启动 DNA 聚合酶可以实现短片段和长片段的同时扩增。使用 Platinum II Taq 热启动 DNA 聚合酶或其它热启动 DNA 聚合酶,在 50μL 反应体系中从 50ng 人基因组 DNA 中扩增 132bp、251bp、1,005bp 和 3.9kb 片段:(A) NEB OneTaq Hot Start DNA Polymerase,(B)Qiagen Fast Cycling PCR Kit,(C)Roche FastStart Taq DNA Polymerase。所有四个目的片段都使用相同的PCR反应条件,图中已标注退火和延伸条件设置。所使用的标准参照物为 Thermo Scientific ZipRuler Express DNA Ladder 2

Platinum II Taq 热启动 DNA 聚合酶是一种经分子改造的酶,DNA 合成速率得到提高。因此,当使用 Platinum II Taq 热启动 DNA 聚合酶时,获得 PCR 结果的速度通常比使用其它热启动 Taq DNA 聚合酶快两倍以上。

Fast cycling reduces PCR run time

图3:快速扩增,缩短PCR运行时间。分别使用 Platinum II Taq 热启动 DNA 聚合酶和来自其它供应商的热启动 DNA 聚合酶,在 50μL 反应体系中,对 50ng 人基因组 DNA 中的 529bp 片段扩增 35 个循环:(A) Sigma- Aldrich KAPA2G Fast HotStart PCR Kit ,(B) NEB OneTaq Hot Start DNA Polymerase,(C) Promega GoTaq G2 DNA Polymerase,(D) Toyobo Quick Taq HS DyeMix,(E) Roche FastStart Taq DNA Polymerase,(F) Sigma-Aldrich JumpStart Taq DNA Polymerase。每种聚合酶的扩增时间以紫色显示,ProFlex PCR系统上的渐变时间(6°C /秒升温速率)以红色显示。在1%TAE琼脂糖凝胶中分析PCR产物,结果显示于图下方。所用的 DNA 标准参照物为 ZipRuler Express DNA Ladder 2

Platinum II Taq 热启动 DNA 聚合酶所具备的高灵敏度,可确保在起始材料有限或样本中目标 DNA 浓度较低的实验中,成功扩增出特定 PCR 产物。 

High sensitivity and reliable amplification from low amounts of input DNA

图4:从少量起始 DNA 中实现高灵敏度和可靠的扩增。使用 Platinum II Taq 热启动 DNA 聚合酶和其它 DNA 聚合酶(A—KAPA2G Fast HotStart,B—NEB OneTaq 热启动,C—Promega GoTaq G2,D—Sigma JumpStart Taq,以及 E—Takara Taq HS Perfect Mix),在 50μL PCR 反应体系中,分别从 0(无模板对照)、0.016、0.08、0.4、2、10、50ng、250ng 人基因组 DNA 中扩增 529bp 片段。预估的拷贝数约为每0.016ng人类基因组DNA 5个拷贝。分子量标准参照物为 ZipRuler Express DNA Ladder 2

Platinum II Taq 热启动 DNA 聚合酶经过分子改造,对抑制剂具有耐受性,可对非最佳纯度的样品实现成功的扩增。

Resistance to inhibitors

图5:耐受抑制剂。使用 Platinum II Taq 热启动 DNA 聚合酶和其它 DNA 聚合酶(A—KAPA 2G Robust HotStart,B—NEB OneTaq 热启动,C—Promega GoTaq G2,以及 D— Takara Taq Hot Start Version),从人基因组 DNA 中扩增 1kb 片段。反应混合物包含:1—腐植酸(终浓度为1.3μg/ mL),2—氯化血红素(终浓度为6μM),3—木聚糖(终浓度为0.26 mg / mL)或4—不含抑制剂对照。分子量标准参照物为 ZipRuler Express DNA Ladder 2

Amplification of DNA extracted from FFPE tissue samples

图6:对从FFPE组织样品中提取的DNA进行PCR扩增。使用 Platinum II Taq 热启动 DNA 聚合酶,从小鼠 FFPE 组织样品中提取的不同量 DNA 中扩增 527bp 片段。使用针对 FFPE 的 RecoverAll 总核酸提取试剂盒提取 DNA。NTC: 无模板对照。PC: 阳性对照(1 ng纯化的小鼠基因组DNA)。分子量标准参照物为 ZipRuler Express DNA Ladder 2

Platinum II Taq 热启动 DNA 聚合酶和 2X 预混液的配方,使得研究人员能够对富含 AT 到富含 GC 的多种靶标进行扩增。产品随附提供一管单独的Platinum GC Enhancer,用于对难以扩增、高GC含量的靶标进行特异性扩增并提高产量。

Robust amplification of AT-rich and GC-rich targets

图7:针对富含AT和GC模板的强劲扩增。在50μL PCR反应体系中,从100ng人类基因组DNA中扩增出GC含量逐渐增加的各种DNA片段(GC含量已在相应泳道上方标注)。Platinum GC Enhancer用于> 65%GC含量的靶标。分子量标准参照物为 ZipRuler Express DNA Ladder 2

由 Platinum II Taq 热启动 DNA 聚合酶产生的 PCR 片段适用于 Sanger 测序。该酶具有出色的性能,通用引物退火温度和快速合成功能,可以轻松简便地生成用于 Sanger 测序的 PCR 扩增子。

高质量的 Sanger 测序结果

图 8.高质量的 Sanger 测序结果。使用 Applied Biosystems 3130xl 遗传分析仪,对由 Platinum II Taq 热启动 DNA 聚合酶扩增的 1.6kb PCR 片段进行 Sanger 测序。显示了内置测序分析软件的 KB basecaller 报告的数据。有效读长(CRL)被定义为给定质量值(QV)下最长的不间断碱基区段。QV20 对应于基本识别错误概率 1%,QV30 对应于 0.1%。QV> 20 被视为高质量,并且在大多数情况下是可接受的。

What is hot-start PCR?

Platinum II Taq 热启动 DNA 聚合酶和 Platinum Taq DNA 聚合酶有何不同?

Platinum Taq 聚合酶是一款历经二十多年一直为研究者所信赖的产品,已被数千种出版物所引用。新一代热启动DNA聚合酶—Platinum II Taq 热启动 DNA 聚合酶是经全新改造、具有扩增快速、强劲性能的聚合酶。我们向启动新项目的研究人员推荐此款产品,产品性能的优越性能使您从中获益。其相关性能总结于下表中。

Platinum II Taq 热启动和 Platinum II Taq DNA 聚合酶技术参数比较

 Platinum II Taq 热启动 DNA 聚合酶Platinum Taq DNA 聚合酶
技术参数
使用通用退火温度
扩增速度15 sec/kb1 min/kb
灵活的延伸步骤a
抑制剂耐受性
目的片段长度长达 5 kb长达 5 kb
热启动修饰抗体介导抗体介导
保真度与 Taq DNA聚合酶比较1x1x
平末端或3‘A端3’A3’A
PCR反应体系的室温稳定性24 h24 h
高GC含量模板
残留的人类和细菌DNAd被控制在极低水平是的
(≤1 拷贝细菌 gDNA/酶单位)
产品形式
预混液无色/绿色b无色/绿色b
单酶无色/绿色c无色/绿色b

a延伸步骤可延长至 60 秒/kb 而不影响扩增的特异性。b选择使用绿色缓冲液,PCR 产物可直接凝胶上样电泳。c绿色缓冲液单独提供,可与单酶配合使用。d在生产 Platinum II Taq 热启动 DNA 聚合酶的过程中,我们采取严格的措施进行控制和验证,以确保每单位聚合酶中含有不超过一个残留细菌基因组 DNA 拷贝。

References

(Micro)biota

UsagePublications
Ion Torrent sequencing of bacterial 16S rRNA genesBasic M, Bolsega S, Smoczek A et al. (2021) Monitoring and contamination incidence of gnotobiotic experiments performed in microisolator cages. Int J Med Microbiol 311(3):151482. 
Metabarcoding of sedimentary ancient DNA samplesCapo E, Giguet-Covex C, Rouillard A et al. (2021) Lake sedimentary DNA research on past terrestrial and aquatic biodiversity: overview and recommendations. Quaternary 4:6. 
Restriction enzyme site-directed amplification PCR (REDA-PCR) and cloning with microalga DNALee JW, Lee MW, Ha JS (2020) Development of a species-specific transformation system using the novel endogenous promoter calreticulin from oleaginous microalgae Ettlia sp. Sci Rep 10(1):13947. 
Diatom DNA amplification with degenerate primers for Illumina sequencingNistal A, Garcia P, Garcia J et al. (2021) DNA metabarcoding and morphological methods show complementary patterns in the metacommunity organization of lentic epiphytic diatoms. ARPHA Conference Abstracts 4: e63672. 
Ion Torrent sequencing of bacterial 16S rRNA genesSchwarz SR, Hirsch S, Hiergeist A et al (2020) Limited antimicrobial efficacy of oral care antiseptics in microcosm biofilms and phenotypic adaptation of bacteria upon repeated exposure. Clin Oral Investig Epub ahead of print 
Amplification of bacterial 16S rRNA gene using barcoded primers for Illumina sequencingWallis A, Yannuzzi IM, Choi MW (2021) Investigating the distribution of strains of Erwinia amylovora and streptomycin resistance in apple orchards in New York using CRISPR profiles: a six-year follow-up. Plant Dis Epub ahead of print. 

Cancer research

UsagePublications
PCR with genomic DNA of primary human cancer cells for mutation detectionAnwar SL, Hasemeier B, Schipper E et al. (2019) LINE-1 hypomethylation in human hepatocellular carcinomas correlates with shorter overall survival and CIMP phenotype. PLoS One 14(5):e0216374. 
PCR with genomic DNA from whole blood samples, followed by Sanger sequencingArévalo-Jaramillo P, Idrobo A, Salcedo L (2019) Biochemical and genotoxic effects in women exposed to pesticides in Southern Ecuador. Environ Sci Pollut Res Int 26(24):24911–24921. 
PCR of bisulfite-treated DNA from carcinoma cell lines, followed by Sanger sequencingIbrahim ML, Klement JD, Lu C et al. (2018) Myeloid-Derived Suppressor Cells Produce IL-10 to Elicit DNMT3b-Dependent IRF8 Silencing to Promote Colitis-Associated Colon Tumorigenesis. Cell Rep 25(11):3036–3046. 
PCR with bisulfite-treated DNA from human tissues and cells, followed by Ion Torrent sequencingMa Y, Chai N, Jiang Q (2020) DNA methyltransferase mediates the hypermethylation of the microRNA 34a promoter and enhances the resistance of patient-derived pancreatic cancer cells to molecular targeting agents. Pharmacol Res 160:105071. 
Touchdown PCR with DNA from FFPE samples, followed by Sanger SequencingMaier AD, Stenman A, Svahn F et al. (2021) TERT promoter mutations in primary and secondary WHO grade III meningioma. Brain Pathol 31(1):61–69. 
Two-step RT-PCR with DNA from human cells for detection of transgene expressionMcCormick CA, Samuels TL, Battle MA (2021) H+/K+ATPase Expression in the Larynx of Laryngopharyngeal Reflux and Laryngeal Cancer Patients. Laryngoscope 131(1):130–135. 
Amplification of an siRNA template from human cancer cell linesWang YL, Chang LC, Chen KB et al. (2021) Aptamer-guided targeting of the intracellular long-noncoding RNA HOTAIR. Am J CancerRes 11(3):945–954. 

(Micro)biota

UsagePublications
Ion Torrent sequencing of bacterial 16S rRNA genesBasic M, Bolsega S, Smoczek A et al. (2021) Monitoring and contamination incidence of gnotobiotic experiments performed in microisolator cages. Int J Med Microbiol 311(3):151482. 
Metabarcoding of sedimentary ancient DNA samplesCapo E, Giguet-Covex C, Rouillard A et al. (2021) Lake sedimentary DNA research on past terrestrial and aquatic biodiversity: overview and recommendations. Quaternary 4:6. 
Restriction enzyme site-directed amplification PCR (REDA-PCR) and cloning with microalga DNALee JW, Lee MW, Ha JS (2020) Development of a species-specific transformation system using the novel endogenous promoter calreticulin from oleaginous microalgae Ettlia sp. Sci Rep 10(1):13947. 
Diatom DNA amplification with degenerate primers for Illumina sequencingNistal A, Garcia P, Garcia J et al. (2021) DNA metabarcoding and morphological methods show complementary patterns in the metacommunity organization of lentic epiphytic diatoms. ARPHA Conference Abstracts 4: e63672. 
Ion Torrent sequencing of bacterial 16S rRNA genesSchwarz SR, Hirsch S, Hiergeist A et al (2020) Limited antimicrobial efficacy of oral care antiseptics in microcosm biofilms and phenotypic adaptation of bacteria upon repeated exposure. Clin Oral Investig Epub ahead of print 
Amplification of bacterial 16S rRNA gene using barcoded primers for Illumina sequencingWallis A, Yannuzzi IM, Choi MW (2021) Investigating the distribution of strains of Erwinia amylovora and streptomycin resistance in apple orchards in New York using CRISPR profiles: a six-year follow-up. Plant Dis Epub ahead of print. 

Cancer research

UsagePublications
PCR with genomic DNA of primary human cancer cells for mutation detectionAnwar SL, Hasemeier B, Schipper E et al. (2019) LINE-1 hypomethylation in human hepatocellular carcinomas correlates with shorter overall survival and CIMP phenotype. PLoS One 14(5):e0216374. 
PCR with genomic DNA from whole blood samples, followed by Sanger sequencingArévalo-Jaramillo P, Idrobo A, Salcedo L (2019) Biochemical and genotoxic effects in women exposed to pesticides in Southern Ecuador. Environ Sci Pollut Res Int 26(24):24911–24921. 
PCR of bisulfite-treated DNA from carcinoma cell lines, followed by Sanger sequencingIbrahim ML, Klement JD, Lu C et al. (2018) Myeloid-Derived Suppressor Cells Produce IL-10 to Elicit DNMT3b-Dependent IRF8 Silencing to Promote Colitis-Associated Colon Tumorigenesis. Cell Rep 25(11):3036–3046. 
PCR with bisulfite-treated DNA from human tissues and cells, followed by Ion Torrent sequencingMa Y, Chai N, Jiang Q (2020) DNA methyltransferase mediates the hypermethylation of the microRNA 34a promoter and enhances the resistance of patient-derived pancreatic cancer cells to molecular targeting agents. Pharmacol Res 160:105071. 
Touchdown PCR with DNA from FFPE samples, followed by Sanger SequencingMaier AD, Stenman A, Svahn F et al. (2021) TERT promoter mutations in primary and secondary WHO grade III meningioma. Brain Pathol 31(1):61–69. 
Two-step RT-PCR with DNA from human cells for detection of transgene expressionMcCormick CA, Samuels TL, Battle MA (2021) H+/K+ATPase Expression in the Larynx of Laryngopharyngeal Reflux and Laryngeal Cancer Patients. Laryngoscope 131(1):130–135. 
Amplification of an siRNA template from human cancer cell linesWang YL, Chang LC, Chen KB et al. (2021) Aptamer-guided targeting of the intracellular long-noncoding RNA HOTAIR. Am J CancerRes 11(3):945–954. 

订购信息

Platinum II Taq 热启动 DNA 聚合酶

Selected references

ApplicationsProduct format usedPublications
Study of genes in cultures of human and mouse pluripotent stem cells—cDNA was amplified by two-step PCR for a number of genes, then visualized with the SYBR Safe gel stain.Platinum II Taq Hot-Start DNA PolymeraseEsseltine JL, Brooks CR, Edwards NA et al. (2020) Dynamic regulation of connexins in stem cell pluripotency. Stem Cells 38:52–66.
Cloning of the sea lamprey GHR gene—GHR was amplified by RT-PCR and TA-cloned.Platinum II Taq Hot-Start DNA Polymerase, supplemented with the included GC EnhancerGong N, Ferreira-Martins D, McCormick SD et al. (2020) Divergent genes encoding the putative receptors for growth hormone and prolactin in sea lamprey display distinct patterns of expression. Sci Rep 10(1):1674.
Amplification of GC-rich (~70%) chromosomal DNA or cDNA— Multiple amplicons were co-cycled following a universal protocol.Platinum II Hot-Start PCR Master Mix, supplemented with the included GC EnhancerHackl T, Martin R, Barenhoff K et al. (2019) Four high-quality draft genome assemblies of the marine heterotrophic nanoflagellate Cafeteria roenbergensis. bioRxiv 751586.
Methylation analysis of genomic DNA from colon cells—Bisulfite-treated DNA was amplified and Sanger sequenced.Platinum II Taq Hot-Start DNA PolymeraseIbrahim ML, Klement JD, Lu C et al. (2018) Myeloid-Derived Suppressor Cells Produce IL-10 to Elicit DNMT3b-Dependent IRF8 Silencing to Promote Colitis-Associated Colon Tumorigenesis. Cell Rep 25(11):3036–3046.
Detection of microRNA‑21 expression levels—qPCR SYBR Green assays were performed using cDNA templates.Platinum II Taq Hot‑Start DNA PolymeraseWei X, You X, Zhang J et al. (2019) miR‑21 inhibitor facilitates the anticancer activity of doxorubicin loaded nanometer in melanoma. Oncol Rep 42(1):414–424.
Amplification of the ITS1 region of Sarcocystis genomic DNA—Sanger sequencing was performed afterward.Platinum II Hot-Start Green PCR Master MixPrakas P, Kirillova V, Gavarāne I et al. (2019) Morphological and molecular description of Sarcocystis ratti n. sp. from the black rat (Rattus rattus) in Latvia. Parasitol Res 118(9):2689–2694.
RT-PCR verification of Sendai reprogramming vector clearance in a transfected iPSC line—Analysis was done on E-Gel agarose gels.Platinum II Hot-Start PCR Master MixHong J, Xu M, Li R et al. (2019) Generation of an induced pluripotent stem cell line (TRNDi008-A) from a Hunter syndrome patient carrying a hemizygous 208insC mutation in the IDS gene. Stem Cell Res 37:101451.

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