RT-PCR & RT-qPCR & RT反轉錄酵素
基因的表達分析,通常從RNA起始。而RNA的多樣性和不穩定性,往往對下有實驗帶來挑戰。Zgenebio的RNA保存/提取產品使用極其方便,避免了繁瑣的液氮凍存、研磨,而獲得的RNA品質同傳統方法一樣好。基於我們專利的HiScript逆轉錄酶衍生的一系列產品,針對RT-PCR和RT-qPCR分別優化;更有簡便的一管式SuperMix提供。
Reverse Transcriptase 反轉錄酶
具有超強耐熱性和延伸效率反轉錄酶
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具有超強耐熱性和延伸效率的新一代反轉錄酶
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• 50℃半衰期超過240分鐘,55℃半衰期可達60分鐘
• 可合成創紀錄的20 kb全長cDNA
• 5分鐘即可合成長達10 kb的cDNA,適合快速反轉錄反應
• 更高的雜質耐受度
• 更高的檢測靈敏度,分子診斷首選反轉錄酶
產品描述
HiScript Reverse Transcriptase的基礎上,我們通過專利的體外分子進化技術,獲得了一個全新的反轉錄酶突變體-HiScript II Reverse Transcriptase。其攜帶的多個點突變使得其熱穩定性大幅提高,在50℃的半衰期超過240分鐘,並可在55℃長時間反應而保持穩定,保證了具有複雜二級結構模版的反轉錄成功率。同時,高溫下保持的高活性能夠帶來更高的cDNA產量。同時,HiScript II Reverse Transcriptase與模版的親和力 (affinity)以及行進性 (processivity)大幅提高,表現為極強的cDNA持續合成能力。HiScript II Reverse Transcriptase可從小鼠骨骼肌總RNA中反轉錄獲得創紀錄的20.0 kb的cDNA (圖2),並且5分鐘內的合成長度可達10 kb,非常適合快速反轉錄反應。HiScript® II Reverse Transcriptase對於常見的反轉錄抑制劑具有更好的耐受度。這些抑制劑通常是RNA純化試劑的組分,或者是來源於樣品,在純化過程中會有不同程度的殘留,並在逆轉錄時被引入,如 guanidine、乙醇、木聚糖、EDTA、SDS、CTAB。與野生型的M-MLV (H-)以及其它反轉錄酶相比,HiScript II Reverse Transcriptase對雜質耐受度更高。這對於來源複雜的樣品 (如FFPE組織),或者富含多糖多酚的植物樣品來說更為重要
• 可合成創紀錄的20 kb全長cDNA
• 5分鐘即可合成長達10 kb的cDNA,適合快速反轉錄反應
• 更高的雜質耐受度
• 更高的檢測靈敏度,分子診斷首選反轉錄酶
產品描述
HiScript Reverse Transcriptase的基礎上,我們通過專利的體外分子進化技術,獲得了一個全新的反轉錄酶突變體-HiScript II Reverse Transcriptase。其攜帶的多個點突變使得其熱穩定性大幅提高,在50℃的半衰期超過240分鐘,並可在55℃長時間反應而保持穩定,保證了具有複雜二級結構模版的反轉錄成功率。同時,高溫下保持的高活性能夠帶來更高的cDNA產量。同時,HiScript II Reverse Transcriptase與模版的親和力 (affinity)以及行進性 (processivity)大幅提高,表現為極強的cDNA持續合成能力。HiScript II Reverse Transcriptase可從小鼠骨骼肌總RNA中反轉錄獲得創紀錄的20.0 kb的cDNA (圖2),並且5分鐘內的合成長度可達10 kb,非常適合快速反轉錄反應。HiScript® II Reverse Transcriptase對於常見的反轉錄抑制劑具有更好的耐受度。這些抑制劑通常是RNA純化試劑的組分,或者是來源於樣品,在純化過程中會有不同程度的殘留,並在逆轉錄時被引入,如 guanidine、乙醇、木聚糖、EDTA、SDS、CTAB。與野生型的M-MLV (H-)以及其它反轉錄酶相比,HiScript II Reverse Transcriptase對雜質耐受度更高。這對於來源複雜的樣品 (如FFPE組織),或者富含多糖多酚的植物樣品來說更為重要
One Step RT-PCR Kit
產品介紹:
One Step RT-PCR Kit專為以RNA為模版(如RNA病毒)的終點法PCR檢測而設計。使用基因專一引物(GSP),逆轉錄和PCR反應在一管內完成,不需要額外的操作,大大提高了檢測速度,並降低了污染的風險。整合Reverse Transcriptase以及AceTaq DNA Polymerase的優越性能,配合經過優化的Buffer ,One Step RT-PCR Kit的檢測靈敏度可達到1 pg總RNA。試劑組以便捷的Master Mix形式提供。2 × Ace-Hi TM Mix包含優化的Buffer和dNTP; Enzyme Mix包含比例優化的Reverse Transcriptase、RNase inhibitor以及AceTaq DNA Polymerase。 ◎方便快速: 使用單管單步驟RT-PCR系統 ◎獨特配方: 使用MMLV RT 及Taq 酵素混合液 ( Enzyme Mix) 以及特殊reaction buffer , 效果穩定. ◎敏感度高: 附獨家配方Thermo-Enhancer, 使RT可在60℃反應, 可解RNA二級結構,提高RT-PCR產量及長度. order info ZGVP601-50 50RXN |
RT-PCR of 220bp zebraf ish EF-1-α gene by RT-PCR kit.
0.5ng zebrafish total RNA was used for RT-PCR, RT step was performed at 50°C and 60°C respectively. Lane 1 : RT-PCR by supplier A Lane 2 : RT-PCR by supplier B with Enhancer Lane 3 : RT-PCR by One RT-PCR kit with Thermo-Enhancer |
One-Step RT-PCR Kit llI (SuperSAMscript III RTase/GenHot)
訂購資訊:ZGGMRP01-III 50RXN
The One-Step RT-PCR Kit llI provides one-tube, two-enzyme system for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The system uses SuperSAMscript III reverse transcriptase for first-strand cDNA synthesis and GenHot DNA polymerase for PCR amplification. SuperSAMscript III RTase is a genetically engineered version of M-MLV RTase which lacks 3’ to 5’ exonucleolytic proofreading function and has a no RNase H activity as compared to AMV RTase, that is thermo stable (amplified DNA in 37~55 ℃ ) and is useful for full-length cDNA amplification(~15kb). GenHot Taq DNA polymerase in the kit is modified with specific monoclonal antibody and can eliminate amplification artifacts such as primer-dimer formation and mis-priming during pre-amplification stage.
Figure 1. Sensitivity of One Step RT-PCR Kit lll.
Amplification of specific RNA from serial dilution of RNA templates with One-Step RT PCR Kit lll. RNA templates were prepared by in vitro transcription, and copies numbers were determined and calculated by spectrophotometer and gel phosphor screen imaging. M: GM100 DNA Ladder Template: E.coli total RNA |
Two-Step SuperSAMscript III RT-PCR Kit
訂購資訊:ZGGMRP012-M3 50RXN
The Two-Step SuperSAMscript III RT-PCR Kit is provided with all components required to perform first-strand cDNA synthesis and second strand DNA amplification. SAMscript RTase is genetically engineered version of M-MLV RTase which is a frequent choice for cDNA synthesis because of its ease of use and low intrinsic RNase H activity. In the cDNA synthesis step, RNA is reverse transcribed by SuperSAMscript III RTase to produce its cDNA. For the subsequent amplification of the cDNA template, PCR Hot-Start Master Mix ll is provided. It can eliminate amplification artifacts such as primer-dimer formation and mis-priming during pre-amplification stage and thus may provide improved specificity. PCR Hot-Start Master Mix II can be used for effective amplification of DNA up to 15 kb in length.
Order info
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