Maxima First Strand cDNA Synthesis Kit for RT-qPCR/第一链cDNA合成试剂盒
货号：K1641,K1642
规格：50 reactions,200 reactions
价格：3606,10692
产品类型：PCR 及DNA聚合酶
品牌：Thermo Fisher
| Thermo Scientific Maxima First Strand cDNA Synthesis Kit for RT-qPCR is a convenient system optimized for cDNA synthesis in 2-step quantitative RT-PCR (RT-qPCR) applications. The kit uses Maxima Reverse Transcriptase (RT), an advanced enzyme derived byin vitroevolution of M-MuLV RT. The enzyme features high thermostability, robustness and increased cDNA synthesis rate compared to wild type M-MuLV RT.The Maxima First Strand cDNA Synthesis Kit for RT-qPCR is capable of reproducible cDNA synthesis from a wide range of total RNA amounts (1 pg to 5 µg) at elevated temperatures (42 to 65°C).The synthesis reaction can be completed in 15 to 30 minutes. Components of the Maxima First Strand cDNA Synthesis Kit for RT-qPCR are pre-mixed to save time and to reduce the possibility of pipetting errors.Additional Information about Reaction ComponentsThe Maxima Enzyme Mix contains Maxima Reverse Transcriptase and Thermo ScientificRiboLock RNase Inhibitor,. The recombinant RiboLock RNase Inhibitor effectively protects RNA templates from degradation by RNases A, B and C at temperatures up to 55°C.5X Reaction Mix contains the remaining reaction components: reaction buffer, dNTPs, oligo(dT)18and random hexamer primers.Water, nuclease-freeis provided for reaction set-up and dilution of sample RNA. The absence of endo-, exodeoxyribonucleases, ribonucleases and phosphatases has been confirmed by appropriate quality tests. |
优点： |
| - High yields of full length cDNA up to 20 kb- Efficient cDNA synthesis at wide temperature range from 42°C to 65°C- Increased synthesis rate—complete cDNA synthesis in 15-30 minutes- High sensitivity and specificity |
数据： |
图1.Effective removal of genomic DNA using dsDNaseTwo-step RT-qPCR analysis of PBGD gene with (RT+) and without (RT-) reverse transcriptase. cDNA synthesis was performed from 0.2ng total Jurkat RNA using Maxima First Strand cDNA Synthesis Kit with dsDNase (#K1671) or other kits including gDNA removal step. Flat RT-plot with Maxima kit indicates complete removal of contaminating gDNA, whereas RT-reactions with other suppliers' kits indicate amplification of residual gDNA.图2.Highly sensitive two step RT-qPCRA – amplification plot.B – standard curve.Amplification of the human PPP1CA gene was performed on varying amounts of Jurkat cell total RNA (5μg, 2.5μg, 1μg, 100ng, 10ng, 1ng, 100pg, 10pg, 1pg). First strand cDNA was generated with the Maxima First Strand cDNA Synthesis Kit for RT-qPCR (#K1641). cDNA was amplified with the Maxima Probe/ROX qPCR Master Mix (2X) using the TaqMan assay specific for PPP1CA. Reactions were performed on an ABI® 7500 Real-Time PCR instrument. 1pg of total RNA was successfully detected. Reaction efficiency was 105%, slope-3.29, R2=0.999.图3.Reproducible RT-qPCR results using Maxima First Strand cDNA Synthesis Kit for RT-qPCRFirst strand cDNA was generated from 100ng to 1pg of total Jurkat cell RNA with the Maxima First Strand cDNA Synthesis Kit for RT-qPCR (#K1641) in 16 replicate reactions. Synthesized cDNA was used as a template in subsequent qPCR with Maxima SYBR Green/ROX qPCR Master Mix (#K0221) on the ABI® 7500 Real-Time PCR instrument. Parallel RT reactions demonstrate reproducible cDNA synthesis and low variability levels with a wide range of starting RNA amounts.图4.Greater yields and higher sensitivity using Maxima First Strand cDNA Synthesis Kit for RT-qPCRAmplification of theE. coli23S RNA gene was performed on 10-fold serial dilutions of total RNA (30ng to 3fg). First strand cDNA was generated using the Maxima First Strand cDNA Synthesis Kit for RT-qPCR (red) and a reverse transcription kit from another vendor (green). cDNA was amplified using Maxima SYBR Green/ROX qPCR Master Mix (#K0221) on the ABI® 7500 Real-Time PCR; instrument. All starting RNA amounts (30ng to 3fg) were successfully detected using the Maxima First Strand cDNA Synthesis Kit with lower Cq and 102% reaction efficiency.图5.dsDNase treatment does not compromise RNA quality and quantitySerial dilutions of total RNA were used in cDNA synthesis with Maxima First Strand cDNA Synthesis kits with and without dsDNase step. No changes in Cq values are observed between the compared protocols. |
组成成分： |
| ▪Maxima Enzyme Mix▪ 5X Reaction Mix▪ nuclease-free water |
相关产品： |
| ▪DNA 片段标准品▪DNA ladder▪DNA 聚合酶▪PCR▪DNA纯化▪RNA纯化 |
技术参数 产品应用 • 2-step RT-PCR
• 2-step RT-qPCR
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