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
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产品简介
本产品是用于去除基因组DNA进行反转录的试剂盒。该试剂盒在42℃，2分钟即可 除去基因组DNA。同时由于反转录试剂中含有抑制gDNA Eraser的组分，经过 gDNA Eraser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，cDNA第一链合成的效率和产 量更高，可利用pg级的总RNA或mRNA合成cDNA第一链。如逆转录产物cDNA用于下 游荧光定量检测，可在42℃，15分钟完成逆转录反应。本试剂盒适用于第一链cDNA 的合成和后续的RT-PCR、RT-qPCR、以及全长cDNA文库的构建等。
产品特点
1．快速去除基因组：含有去除基因组DNA的gDNA Eraser，只需2分钟即可除去基因 组DNA。
2．快速逆转录：15分钟即可获得荧光定量PCR模板cDNA第一链合成。
3．灵敏度高：可利用pg级总RNA或mRNA模板合成cDNA第一链。
4．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．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。
3．反应体系可倍比放大，10 μl反应体系可最大使用1 μg总RNA。
4．Primer Mix由Oligo(dT)和Random primer配制而成， 也可根据实验需要选用 Oligo-dT Primer或Gene Specific Primer。
5．若起始RNA的量小于50 ng，建议加入RNA酶抑制剂（RNasin）。
6．对于二级结构复杂的RNA模板，建议在操作步骤之前，将模板RNA在65℃孵育5分 钟立刻置于冰上，短暂离心后进行下一步操作。
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。
注意：1） 如果总RNA量大于1 µg，请按比例扩大反应体系。若起始RNA的量小于50 ng，则建议加入RNA酶抑制剂（RNasin）。
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。
注意：1） 可根据实验需要可使用Oligo-dT Primer或Gene Specific Primer，建议20 μl 反应体系 Oligo-dT Primer 50pmol，或Gene Specific Primer 2 pmol。
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：
1）若下游进行荧光定量PCR检测，42℃孵育15分钟，85℃孵育5分钟。
2）若下游进行普通PCR检测，42℃孵育30-50分钟，85℃孵育5分钟。 注意：对于二级结构复杂或GC含量高的模板，可以提高逆转录温度至50℃，增强逆转录效率。
4．反应结束后，短暂离心后置于冰上，再进行后续PCR或荧光定量PCR，如果需要长时 间保存，请置于-20℃。
注意：进行Real-time PCR反应时，逆转录产物的加量应不超过PCR反应总体积的1/10。
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Product Introduction
This product is a kit for removing genomic DNA for reverse transcription. The kit removes genomic DNA in 2 minutes at 42°C. Since the reverse transcription reagent contains a component that inhibits gDNA Eraser, cDNA can be synthesized directly by reverse transcription of gDNA Eraser-treated samples.The kit is equipped with a new high-efficiency reverse transcription enzyme, HiFiScript, with novel mutation sites that dramatically increase the transcriptional activity of the enzyme, resulting in higher efficiency and yield of cDNA first-strand synthesis. The first strand of cDNA can be synthesized with higher efficiency and yield, and the first strand of cDNA can be synthesized from pg total RNA or mRNA. If the reverse transcription product cDNA is used for downstream fluorescence quantitative detection, the reverse transcription reaction can be completed at 42℃ in 15 minutes. This kit is suitable for the synthesis of first-strand cDNA and subsequent RT-PCR, RT-qPCR, and the construction of full-length cDNA libraries.
Product Features
1. Rapid genome removal: contains gDNA Eraser for genomic DNA removal, which removes genomic DNA in just 2 minutes.
2. Rapid reverse transcription: 15 minutes to obtain fluorescent quantitative PCR template cDNA first strand synthesis.
3. High sensitivity: cDNA first strand can be synthesized using pg-level total RNA or mRNA templates.
4. Highly efficient reverse transcription: Novel mutation sites dramatically increase enzyme activity, resulting in higher yields of cDNA.
matters needing attention
1. During operation, RNase contamination should be avoided to prevent RNA degradation or cross-contamination in the experiment. It is recommended that operators wear masks and disposable gloves and change the gloves frequently, and use specialized instruments and consumables.
2. The reverse transcription system is prepared and operated on ice to prevent degradation of RNA. Store the kit enzymes at -20ºC as soon as possible after use and try to avoid repeated freezing and thawing.
3. The reaction system can be scaled up to a maximum of 1 μg of total RNA in 10 μl of reaction system.
4. Primer Mix is prepared by Oligo(dT) and Random primer, and Oligo-dT Primer or Gene Specific Primer can also be used according to the experimental needs.
5. If the amount of starting RNA is less than 50ng, it is recommended to add RNAase inhibitor (RNasin).
6. For RNA templates with complex secondary structures, it is recommended to incubate the template RNA at 65°C for 5 minutes immediately on ice prior to the manipulation step and centrifuge briefly before proceeding to the next step.
Usage
Thaw template RNA on ice; place kit components on ice immediately after thawing at room temperature. Each solution was mixed by vortexing and shaking before use and briefly centrifuged.
I. Genomic DNA removal reactions
1. Prepare the reaction system according to the following table on ice in a total volume of 10 μl. To ensure the accuracy of the reaction solution preparation, prepare the premixed system in the amount of reaction number + 2 before dispensing it into each reaction tube and finally adding the RNA sample.
Note: 1) If the amount of total RNA is greater than 1µg, scale up the reaction system proportionally. If the amount of starting RNA is less than 50ng, it is recommended to add RNAase inhibitor (RNasin).
2. Mix by vortex shaking and centrifuge briefly so that the solution on the walls of the tube collects at the bottom.
3. Incubate at 42°C for 2 minutes (this can be extended to 30 minutes for room temperature reactions).
4．At the end of the reaction, centrifuge briefly and place on ice to cool.
II. Reverse transcription reaction
1. Prepare the reaction system on ice according to the following table. In order to ensure the accuracy of the reaction solution configuration, first prepare a premixed solution in the amount of number + 2, and then dispense 10 μl into each reaction tube, take 10 μl of the prepared premixed solution and add it to the reaction tube of step 1 where the de-etching of the genome has been completed.
Note: 1) Oligo-dT Primer or Gene Specific Primer can be used according to the needs of the experiment, it is recommended to use 50 pmol of Oligo-dT Primer or 2 pmol of Gene Specific Primer for 20 μl reaction system.
2. Mix well and centrifuge briefly so that the solution on the walls of the tube collects at the bottom.
3. cDNA synthesis reaction conditions:
1) If fluorescent quantitative PCR assay is performed downstream, incubate at 42°C for 15 minutes and 85°C for 5 minutes.
2) If downstream for normal PCR assay, incubate at 42°C for 30-50 minutes and 85°C for 5 minutes. Note: For templates with complex secondary structure or high GC content, the reverse transcription temperature can be increased to 50°C to enhance reverse transcription efficiency.
4. At the end of the reaction, centrifuge briefly and place on ice before proceeding with subsequent PCR or fluorescence quantitative PCR, or place at -20°C if prolonged storage is required.
Note: When performing Real-time PCR reactions, the amount of reverse transcription product added should not exceed 1/10 of the total volume of the PCR reaction.
规格
别名
HiFiScript gDNA 去除 cDNA 合成试剂盒
英文名称
HiFiScript gDNA Removal cDNA Synthesis Kit
储存条件
-20°C储存,避免反复冻融
运输条件
超低温运输
产品线
生物产品
名称和识别符
分子类型
试剂盒
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
常见问题
本产品应如何储存？
请于?20 °C条件下保存。避免反复冻融。建议按单次用量分装保存。
本产品如何运输？
本产品采用保温箱配冰袋运输。收货后请立即拆包，并按上述储存条件保存。
如何获取批次专用COA？
每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
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