计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
-20°C储存,避免反复冻融。超低温运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
本产品是用于去除基因组DNA进行逆转录的试剂盒，在42℃，2分钟即可除去基因 组DNA。同时，逆转录试剂中含有抑制gDNA Remover的组分，经过gDNA Remover 处理后的样品可以直接进行逆转录反应合成cDNA。
本试剂盒配有新型高效反转录酶HiFiScript，新颖突变位点大幅提升酶的转录活 性。同时，逆转录反应只需15分钟即可完成cDNA第一链的合成。5×HifiScript RT MasterMix为逆转录预混液，包含逆转录所需全部试剂，操作方便快捷。
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产品特点
1．快速去除基因组：含有去除基因组DNA的gDNA Remover，只需2分钟即可除去基因组DNA。
2．快速逆转录：15分钟即可完成cDNA第一链合成。
3．方便快捷：即用型逆转录Mix，操作简便。
4．灵敏度高：可利用pg级总RNA或mRNA模板合成cDNA第一链。
5．高效的逆转录效率：新颖突变位点大幅提升酶活性能，获得更高产量的cDNA。
注意事项
1．在操作过程中应避免RNase污染，防止RNA降解或实验中的交叉污染，建议操作人员带口罩和一次性手套并经常更换手套，使用专门的仪器和耗材。
2．实验尽量使用一次性塑料器皿，若使用玻璃器皿，应使用0.1％DEPC（焦碳酸二乙 1．酯）水溶液在37℃处理12小时，并在120℃下高压灭菌30分钟后使用，或者将玻璃器皿在180℃下干热灭菌60分钟后使用。实验中用到的无菌水应使用0.1%的DEPC处理后进行高压灭菌。
3．逆转录体系配制在冰上进行操作，防止RNA发生降解。试剂盒的酶使用后尽快置于-20 ºC保存，并尽量避免反复冻融。
使用方法
将模板RNA在冰上解冻；试剂盒组分在室温解冻后立刻置于冰上。使用前将每种 溶液涡旋振荡混匀，并经短暂离心后使用。
一、去除基因组DNA反应
1．根据以下表格在冰上配制反应体系，总体积为10 μl。为了保证反应液配制的准确性， 先按反应数+2的量配制预混体系， 然后再分装到每个反应管中， 最后加入RNA样品。
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注意：1） 如果总RNA量大于1 µg，请按比例扩大反应体系。
2．涡旋震荡混匀，短暂离心，使管壁上的溶液收集到管底。
3．42℃孵育2分钟（室温反应时，可以延长到30分钟）。
4．反应结束后，短暂离心，置于冰上冷却。
二、逆转录反应
1．根据以下表格在冰上配制反应体系，反应液配制请在冰上进行。为了保证反应液配置的准确性，先按反应数+2的量配制成预混溶液，然后再分装10 μl到每个反应管中，取配制的预混液10 μl加入至已完成去基因组的步骤1反应管中。
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2．混匀，短暂离心，使管壁上的溶液收集到管底。
3．cDNA合成反应条件：37℃孵育15分钟，85℃孵育5秒钟。
4．反应结束后，短暂离心后置于冰上，再进行后续反应，如果需要长时间保存，请置于-20℃。
This product is a reagent kit for reverse transcription of genomic DNA, which can remove genomic DNA in 2 minutes at 42 ℃. Meanwhile, the reverse transcription reagent contains components that inhibit gDNA Remover, and the samples treated with gDNA Remover can be directly subjected to reverse transcription reaction to synthesize cDNA.
This reagent kit is equipped with a novel and efficient reverse transcriptase HiFiScript, which significantly enhances the transcriptional activity of the enzyme through novel mutation sites. Meanwhile, reverse transcription reaction only takes 15 minutes to complete the synthesis of the first strand of cDNA. 5 × HifiScript RT MasterMix is a reverse transcription premix that contains all the reagents required for reverse transcription, making it easy and fast to operate.
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Product features
1. Quick genome removal: gDNA Remover containing genomic DNA can remove genomic DNA in just 2 minutes.
2. Rapid reverse transcription: cDNA first strand synthesis can be completed in 15 minutes.
3. Convenient and fast: Ready to use reverse transcription Mix, easy to operate.
4. High sensitivity: cDNA first strand can be synthesized using pg level total RNA or mRNA templates.
5. Efficient reverse transcription efficiency: Novel mutation sites significantly enhance enzyme activity performance, resulting in higher yields of cDNA.
Matters needing attention
1. During the operation, RNase contamination should be avoided to prevent RNA degradation or cross contamination during experiments. It is recommended that operators wear masks and disposable gloves, frequently change gloves, and use specialized instruments and consumables.
2. Disposable plastic containers should be used as much as possible for experiments. If glassware is used, a 0.1% DEPC (diethyl carbonate 1. ester) aqueous solution should be treated at 37 ℃ for 12 hours, and then sterilized under high pressure at 120 ℃ for 30 minutes before use. Alternatively, glassware should be sterilized under dry heat at 180 ℃ for 60 minutes before use. The sterile water used in the experiment should be treated with 0.1% DEPC and then subjected to high-pressure sterilization.
3. The reverse transcription system is prepared on ice for operation to prevent RNA degradation. The enzyme in the reagent kit should be stored at -20 º C as soon as possible after use, and repeated freeze-thaw should be avoided as much as possible.
Usage
Thaw the template RNA on ice; After thawing at room temperature, immediately place the components of the reagent kit on ice. Before use, mix each solution by vortex oscillation and centrifuge briefly before use.
1、 Genomic DNA removal reaction
1. Prepare a reaction system on ice according to the following table, with a total volume of 10 μ L. To ensure the accuracy of the preparation of the reaction solution, a pre mixed system is first prepared in the amount of reaction number+2, then divided into each reaction tube, and finally RNA samples are added.
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Note: 1) If the total RNA amount is greater than 1 µ g, please expand the reaction system proportionally.
2. Vortex shake and mix well, briefly centrifuge to collect the solution on the pipe wall to the bottom of the pipe.
3. Incubate at 42 ℃ for 2 minutes (during room temperature reaction, it can be extended to 30 minutes).
4. After the reaction is complete, centrifuge briefly and cool on ice.
2、 Reverse transcription reaction
1. Prepare the reaction system on ice according to the following table, and prepare the reaction solution on ice. To ensure the accuracy of the reaction solution configuration, first prepare a premixed solution in the amount of reaction number+2, and then divide it into 10 batches μ Take 10% of the prepared premix from each reaction tube μ Add to the reaction tube of step 1 that has completed genome removal.
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2. Mix well and centrifuge briefly to collect the solution on the tube wall to the bottom of the tube.
3. cDNA synthesis reaction conditions: Incubate at 37 ℃ for 15 minutes and 85 ℃ for 5 seconds.
4. After the reaction is completed, centrifuge briefly and place it on ice before proceeding with the subsequent reaction. If it needs to be stored for a long time, please place it at -20 ℃.
规格
英文名称
HiFiScript gDNA Removal RT MasterMix
储存条件
-20°C储存,避免反复冻融
运输条件
超低温运输
名称和识别符
分子类型
小分子
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
技术文档和文章
溶液计算器
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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