Platinum II Taq Hot-Start DNA Polymerase

專為高特異性和高產量所打造的酶，並允許快速循環

Invitrogen Platinum II Taq熱啟動DNA聚合酶旨在幫助您更快地達到研究目的。透過通用引子黏合功能，可減少反應優化步驟，並允許於co-cycling進行不同的PCR檢測。整合了創新型緩衝液、高性能Taq DNA聚合酶和卓越的熱啟動技術的獨特組合，即使在最嚴苛的實驗應用中，也能獲得出色的PCR結果。

立即訂購 申請樣品 相關資源

產品亮點

  • 通用的引子黏合溫度（60°C）—簡化了引子黏合的優化步驟；透過在共循環中進行不同PCR擴增反應，有助於省去重複多次的PCR運行。
  • DNA合成速度提高4倍，更高的抑制劑耐受性—採用極其穩定的工程改造Taq聚合酶。
  • Platinum熱啟動技術—增強了PCR的特異性、靈敏度及產量；並允許於室溫條件下進行反應設置。
  • 綠色緩衝液規格—透過PCR產物可直接於凝膠上樣，有助於減少移液錯誤。

專為更好的PCR擴增而設計

點擊以下按鈕，了解Platinum II Taq熱啟動DNA聚合酶的優勢。

 

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影片：關於這種酶的優勢

Platinum II Taq熱啟動DNA聚合酶的優勢

通用黏合溫度的擴增方案

使用常規PCR試劑進行PCR檢測時，由於不同的引子黏合溫度和延伸步驟的持續時間不一致，需要針對每個DNA片段設計特定的PCR擴增方案。因此，不同的PCR檢測不能同時在一次PCR運行中進行擴增。透過Platinum II Taq熱啟動DNA聚合酶，可以使用同一個實驗方案和通用的引子黏合溫度，且依據擴增的最長片段來決定延伸步驟，因而可實現同時進行不同的PCR反應。此外，Platinum II Taq熱啟動DNA聚合酶是一種"快速"的DNA聚合酶；因此，這種次世代DNA聚合酶結合通用擴增方案後，透過快速擴增循環，實現短短30分鐘以內完成所有的PCR反應。

Platinum2TaqHot-StartDNA-Polymerase-cycle-together

圖1. 可同時進行PCR擴增，有效的節省時間。使用常規PCR試劑進行PCR擴增時，由於不同引子的黏合溫度和延伸步驟不一致，所以需要針對每個DNA片段設計特定的擴增方案。因此，使用常規PCR試劑時，往往無法在同一次PCR運行中同時擴增多個目標片段。使用Platinum II Taq熱啟動DNA聚合酶，其具有通用的引子黏合溫度，可針對最長片段選擇擴增步驟時間，以實現在一個方案中同時進行不同的PCR擴增。此外，Platinum II Taq熱啟動DNA聚合酶是一種快速DNA聚合酶，僅需30分鐘便可獲得PCR結果。

Time saving enabled by assay co-cycling

圖2. Platinum II Taq熱啟動DNA聚合酶可實現短片段和長片段的同時擴增。使用Platinum II Taq熱啟動DNA聚合酶或其他熱啟動DNA聚合酶，在50 μL反應中從50 ng人類基因組DNA擴增132 bp、251 bp、1,005 bp和3.9 kb片段：(A) NEB OneTaq Hot Start DNA聚合酶，(B) Qiagen快速循環PCR試劑盒，(C) Roche FastStart Taq DNA聚合酶。所有四個目標物片段均使用相同的PCR方案，黏合溫度和延伸條件設置如圖中所示。所使用的標準參照物為Thermo Scientific ZipRuler Express DNA Ladder 2。

快速循環

Platinum II Taq熱啟動DNA聚合酶是一種經工程改造的酶，可提高DNA的合成速率。因此，使用Platinum II Taq熱啟動DNA聚合酶與其他熱啟動Taq DNA聚合酶相比，獲得PCR結果的速度通常會加快兩倍以上。

Fast cycling reduces PCR run time

圖3. 快速循環可縮短PCR的運行時間。分別使用Platinum II Taq熱啟動DNA聚合酶和其他供應商的熱啟動DNA聚合酶，在50 μL反應中，對50 ng人類基因組DNA的529 bp片段進行35個循環擴增：(A) Sigma-Aldrich KAPA2G Fast HotStart PCR試劑盒、(B) NEB OneTaq熱啟動DNA聚合酶、(C) Promega GoTaq G2 DNA聚合酶、(D) Toyobo Quick Taq HS DyeMix、(E) Roche FastStart Taq DNA聚合酶，和 (F) Sigma-Aldrich JumpStart Taq DNA聚合酶。每種聚合酶的擴增時間以紫色顯示，而ProFlex PCR系統的漸變時間（模塊升溫速率為6°C/秒）以紅色顯示。在1% TAE瓊脂糖凝膠中分析PCR產物，結果顯示如圖下方。所使用的標準參照物為ZipRuler Express DNA Ladder 2。

高靈敏度和特異性

Platinum II Taq熱啟動DNA聚合酶所具備的高靈敏度，可確保在起始材料數量有限或樣品中目標DNA濃度較低的實驗中，成功擴增出特定的PCR產物。

High sensitivity and reliable amplification from low amounts of input DNA

圖4. 從少量起始DNA中，實現高靈敏度和可靠的擴增。使用Platinum II Taq Hot-Start DNA聚合酶或其他供應商DNA聚合酶（A—KAPA2G Fast HotStart，B—NEB OneTaq Hot Start，C—Promega GoTaq G2，D—Sigma JumpStart Taq，和E—Takara Taq HS Perfect Mix），在50 μL PCR反應中，分別從0 ng（無模板對照）、0.016 ng、0.08 ng、0.4 ng、2 ng、10 ng、50 ng、250 ng人類基因組DNA中擴增529 bp片段。預估的複製數，每0.016 ng人類基因組DNA約為5個拷貝。使用分子量標準參照物為ZipRuler Express DNA Ladder 2。

對於抑制劑具有耐受性

Platinum II Taq熱啟動DNA聚合酶經過分子改造設計，對抑制劑具有耐受性，可幫助純度不理想的樣品實現成功的擴增。

Resistance to inhibitors

圖5. 對抑制劑的耐受性。使用Platinum II Taq熱啟動DNA聚合酶或其他供應商DNA聚合酶（A—KAPA 2G Robust HotStart，B—NEB OneTaq Hot Start，C—Promega GoTaq G2，和D—Takara Taq Hot Start Version），從人類基因組DNA中擴增1 kb片段。反應混合物包含：1—腐植酸（最終濃度為1.3 µg/mL），2—氯化血紅素（最終濃度為6 µM），3—木聚醣（最終濃度為0.26 mg/mL），或4—無抑制劑對照組。使用的分子量標準參照物為ZipRuler Express DNA Ladder 2。

Amplification of DNA extracted from FFPE tissue samples

圖6. 對從FFPE組織樣品中提取的DNA，進行PCR擴增。使用Platinum II Taq熱啟動DNA聚合酶，從小鼠FFPE組織樣品中提取不同總量的DNA中擴增527 bp片段。使用適用於FFPE的RecoverAll總核酸分離試劑盒RecoverAll總核酸分離試劑盒，進行DNA的提取。NTC：無模板對照組。PC：陽性對照組，取自1 ng純化的小鼠基因組DNA。使用的分子量標準參照物為ZipRuler Express DNA Ladder 2。

穩健的擴增

Platinum II Taq熱啟動DNA聚合酶和2X預混液的配方，可實現對於各式多樣目標物的擴增，涵蓋了從富含AT到富含GC的標的。產品隨附提供一管單獨的Platinum GC Enhancer，可對於高GC含量的目標物進行特異性擴增並提高產量。

Robust amplification of AT-rich and GC-rich targets

圖7. 針對富含AT和GC目標物的穩健擴增。在50 µL PCR反應系統中，從100 ng人類基因組DNA擴增出GC含量逐漸增加的各種DNA片段（GC含量標註於相對應的條帶上方）。Platinum GC Enhancer適用於GC含量大於65%的目標物。所使用的分子量標準參照物為ZipRuler Express DNA Ladder 2。

與Sanger定序法的兼容性

由Platinum II Taq熱啟動DNA聚合酶生成的PCR片段非常適用於Sanger定序。由於該酶的卓越性能、通用型引子黏合溫度和快速合成特性，可以輕鬆簡單地生成適用於Sanger定序的PCR擴增子。

High-quality Sanger sequencing results

圖8. 高品質的Sanger定序結果。使用Applied Biosystems 3130 xl基因分析儀，對由Platinum II Taq熱啟動DNA聚合酶擴增的1.6 kb PCR片段進行了Sanger定序。圖顯示為KB basecaller內建定序分析軟體的所提供之報告數據。有效範圍讀取長度（Clear-range read length，CRL）的定義為給定質量值（Quality Value，QV）下最長的不間斷鹼基片段。QV20相當於1%的基本識別錯誤概率，QV30相當於0.1%的基本識別錯誤概率。QV>20被視為高品質結果，並且在大多數情況下是可接受的。

什麼是熱啟動PCR？

了解PCR擴增中的常見問題，以及如何透過熱啟動PCR解決這些問題。您還將了解不同類型的熱啟動酶修飾，以及如何選擇適合您PCR的熱啟動DNA聚合酶。

Platinum II Taq熱啟動DNA聚合酶和Platinum Taq DNA聚合酶有何不同？

Platinum Taq DNA聚合酶二十多年來一直深受研究人員的信賴，並已被數千篇期刊論文所引用。次世代熱啟動DNA聚合酶—Platinum II Taq熱啟動DNA聚合酶經過全新改造，具有擴增快速、穩健的性能。我們向啟動新項目的研究人員推薦此款Platinum II Taq DNA聚合酶，以便感受卓越的性能並從中獲益，其相關優勢總結如下表。

Platinum II Taq熱啟動DNA聚合酶和Platinum Taq DNA聚合酶的技術參數比較

 Platinum II Taq熱啟動DNA聚合酶Platinum Taq DNA聚合酶
通用型黏合擴增方案是否
擴增速度15秒/kb1分鐘/kb
靈活的擴展步驟*是否
對於抑制劑的耐受性是否
目標物片段長度長達5 kb長達5 kb
熱啟動修飾抗體調控抗體調控
保真度vs. Taq DNA聚合酶1x1x
擴增子突出端類型3’A3’A
PCR反應設置的穩定性24小時24小時
是否可擴增富含GC的模板是是
經認證的人類和細菌DNA低殘留含量d是
（≤1 copy細菌gDNA/酶單位）		否
預混液規格無色/綠b無色/綠b
Stand-alone enzyme formats無色c無色/綠b

a延伸步驟可延長至60秒/kb，而不影響擴增的特異性。b選擇使用綠色緩衝液選項，PCR產物可直接進行凝膠上樣。c綠色緩衝液作為單獨產品提供，可與單酶搭配使用於直接凝膠上樣。d在Platinum II Taq熱啟動DNA聚合酶的生產過程中，我們採取了嚴格的措施控制並透過qPCR驗證，以確保每單位聚合酶中殘留的細菌基因組DNA不超過一個拷貝數。

參考文獻

(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 Cancer Res 11(3):945–954. 
(Micro)biota

(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

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 Cancer Res 11(3):945–954. 
Developmental
biology

Developmental biology

UsagePublications
Two-step RT-PCR with human and mouse pluripotent stem cellsEsseltine JL, Brooks CR, Edwards NA et al. (2020) Dynamic regulation of connexins in stem cell pluripotency. Stem Cells 38:52–66. 
MicroRNA-binding site cloning of bovine DNAShen X, Tang J, Ru W et al. (2021) CircINSR regulates fetal bovine muscle and fat development. Front Cell Dev Bio 8:615638. 
Disease
model

Disease models

UsagePublications
RT-PCR with human iPSC cDNAHong 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. 
Two-step RT-PCR with mouse liver cDNA for alternative splicing assaysHyun J, Sun Z, Ahmadi AR, Bangru S et al. (2020) Epithelial splicing regulatory protein 2-mediated alternative splicing reprograms hepatocytes in severe alcoholic hepatitis. J Clin Invest 130(4):2129–2145. 
PCR of protozoan genomic DNA, followed by Sanger sequencingPrakas 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. 
Mouse genotyping PCRSava V, Fihurka O, Khvorova A et al. (2020) Enriched chitosan nanoparticles loaded with siRNA are effective in lowering Huntington's disease gene expression following intranasal administration. Nanomedicine 24:102119. 
Gene
regulation

Gene regulation

UsagePublications
GC-rich PCR of sea lamprey gene, followed by cloning and sequencingGong 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. 
Two-step RT-PCR with cDNA from goat primary cells for gene expression analysisLuan Z, Fan X, Song H et al. (2019) Testosterone promotes GPX5 expression of goat epididymal epithelial cells cultured in vitro. In Vitro Cell Dev Biol Anim 55(9):677–685. 
Chromatin immunoprecipitation (ChIP) assaysSun D. (2019) Chromatin Immunoprecipitation Assay to Analyze the Effect of G-Quadruplex Interactive Agents on the Binding of RNA Polymerase II and Transcription Factors to a Target Promoter Region. Methods Mol Biol 2035:233–242. 
Phylogenomics

Phylogenomics

UsagePublications
GC-rich PCR of flagellate DNAHackl T, Martin R, Barenhoff K et al. (2020) Four high-quality draft genome assemblies of the marine heterotrophic nanoflagellate Cafeteria roenbergensis. Sci Data 7(1):29. 
Subcloning of E. coli genomic DNA fragmentPrahlad J, Yuan Y, Lin J et al. (2020) The DUF328 family member YaaA is a DNA-binding protein with a novel fold. J Biol Chem 295(41):14236–14247. 
PCR of mitochondrial DNA from blood samplesWharton D, Morey KC, Hanner R. (2021) Maternal inheritance of mitochondrial DNA in mice after inter-species hybridization and 138 generations of backcrossing. Mitochondrial DNA A DNA Mapp Seq Anal 32(2):73–75. 
Plant
biology

Plant biology

UsagePublications
Plant DNA cloningArrey-Salas O, Caris-Maldonado JC, Hernández-Rojas B (2021) Comprehensive genome-wide exploration of C2H2 zinc finger family in grapevine (Vitis vinifera L.): Insights into the roles in the pollen development regulation. Genes (Basel) 12(2):302. 
SNP genotyping of plant by ddRAD-SeqYamazaki A, Shirasawa K, Hosokawa M (2020) Transgressive segregation and gene regions controlling thermotolerance of fruit set and pollen germination in Capsicum chinense. Euphytica 216:179. 
Viral
research

Viral research

UsagePublications
PCR with DNA from FFPE samples for detection of viral (EBV, HPV) sequencesGupta I, Al Farsi H, Jabeen A et al. (2020) High-Risk Human Papillomaviruses and Epstein-Barr Virus in Colorectal Cancer and Their Association with Clinicopathological Status. Pathogens 9(6):452. 
Amplification of fowl adenoviral DNA from allantoic samples, followed by Sanger sequencingWibowo MH, Sahesty A, Mahardika BK et al. (2019) Epizootiology, Clinical Signs, and Phylogenetic Analysis of Fowl Adenovirus in Chicken Farms in Indonesia from 2018 to 2019. Avian Dis 63(4):619–624. 

訂購資訊

Platinum II Taq熱啟動DNA聚合酶

The original Platinum Taq DNA Polymerase
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