计算溶液所需的质量、体积或浓度。
通用型植物总RNA快速提取试剂盒
有货
为什么选择此级别
BioReagent,分子生物学级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
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室温。常规运输 。请查阅批次 COA 获取详细规格。
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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
本试剂盒采用了经过优化的缓冲液系统，可以对各种植物组织，尤其是多糖多酚植物组织进行总RNA的提取，适用范围广泛，提取过程中无需额外使用苯酚、氯仿或巯基乙醇等有毒有害试剂。
本试剂盒具有安全、高效、快速、方便的特点，利用该试剂盒提取的RNA纯度高，极少含蛋白质、基因组DNA和其他杂质的污染。使用本试剂盒可以从100-150 mg植物组织中纯化得到多至几十微克的高纯度RNA。提取得到的RNA可以直接用于Northern杂交、斑点杂交、mRNA纯化、体外翻译、RNA分解酶的保护分析、RT-PCR、Real Time RT-PCR和构建cDNA文库等各种分子生物学实验。
注意事项
1. 操作人员应戴一次性口罩和手套，实验过程中要勤换手套，并使用无RNase的塑料制品及枪头，避免RNase污染。
2. 本试剂盒可除去大部分基因组DNA污染，无需DNase I消化即可用于下游实验，但不同的样本或样本的不同投入量核酸含量差异较大，若下游实验对RNA纯度要求比较严格，可以选择性地进行DNase I消化，参见步骤5，DNase I可单独进行购买。
3. 推荐使用新鲜植物样本进行提取，若无法及时提取，样本应液氮冷冻后冻存于-70℃以下。同时应避免反复冻融，否则影响RNA提取得率和质量。
操作步骤
使用前准备：在 Buffer RW2 中加入48 mL的无水乙醇。
1. 匀浆处理
在2 mL离心管中加入600 μL Buffer RCL，将植物叶片或果实果肉在液氮中迅速研磨成粉末，称取50-100 mg样本至含有Buffer RCL的离心管中，盖上管盖，上下剧烈摇晃20次左右使样本与RCL混合。加入10 μL Proteinase K，立即涡旋振荡30 sec，14,000 rpm (18,630×g) 离心2 min。
注意：
如果样本RNA含量低，可以加入800 μL Buffer RCL，同时将样本量增加到100-130 mg。
如果提取干种子样本，需要使用前在Buffer RCL中加入RE Reagent（可免费提供）。
2. 转移≤ 700 μL的上清液（不同样本的上清液体积不同）至基因组清除柱DE（清除柱放在收集管中），14,000 rpm (18,630 × g) 离心30 sec，弃去基因组清除柱，保留收集管中的滤液。
3. 用移液器量下滤液体积（不同样本的滤液体积会有差异，同一类型同一部位的组织样本只
需量取一个即可），向滤液中加入0.7倍滤液体积的Buffer RBE（此时可能会出现沉淀或絮状 物，属正常现象），混匀后将混合液转移至吸附柱RA（吸附柱放在收集管中），每次上样量≤ 700 μL（如果混合液体积超过700 μL，需要分两次转入），14,000 rpm (18,630×g) 离心30 sec，弃废液，将吸附柱放回收集管中。
4. 如果不进行DNase I消化，向吸附柱中加入700 μL Buffer RW，12,000 rpm (13,680×g) 离心 30 sec，弃废液，将吸附柱放回收集管中，接着按照步骤6-9进行。
5. DNase I 消化（可选）
若后续实验对RNA纯度要求比较严格，可以选择性地进行DNase I 消化。若选择DNase I 消化则跳过步骤4，直接按照步骤5-9进行。
5.1 向吸附柱中加入350 μL Buffer RW，12,000 rpm (13,680×g) 离心30 sec，弃废液，将吸附柱放回收集管中。
5.2 DNase I 储存液的配制：将DNase I 干粉 (2000U) 溶解在550 μL RNase-Free ddH₂O中，轻柔混匀，分装后-30~-15℃贮存（可保存9个月）。
注意：从-30~-15℃融化后的DNase I 储存液保存于2-8℃（可保存6周），不要再次冻存。
5.3 DNase I 工作液的配制：取10 μL DNase I 储存液放入新的RNase-Free离心管中，加入70 μL DNase I Buffer，轻柔混匀。
5.4 向吸附柱中央加入80 μL的DNase I 工作液，室温放置15 min。
5.5 向吸附柱中加入350 μL Buffer RW，12,000 rpm (13,680×g) 离心30 sec，弃废液，将吸附柱放回收集管中。
6. 向吸附柱中加入500 μL Buffer RCW，12,000 rpm (13,680×g) 离心30 sec，弃废液，将吸附柱放回收集管中。
7. 向吸附柱中加入700 μL Buffer RW2（使用前请先加入无水乙醇），12,000 rpm (13,680×g) 离 心30 sec，弃废液，将吸附柱放回收集管中。
8. 12,000 rpm (13,680×g) 离心2 min。
9. 将吸附柱转移至一个新的1.5 mL RNase-Free离心管中，打开吸附柱管盖，室温放置1 min，以彻底晾干吸附柱中的无水乙醇。向吸附膜中间部位悬空滴加30-100 μL RNase-Free ddH₂O，室温放置1 min，12,000 rpm (13,680×g) 离心30 sec。得到的RNA可直接用于下游实验，或于-80℃以下保存。
注意：RNase-Free ddH₂O 体积不应小于30 μL，体积过小影响回收率。若要提高RNA的产量，可将第一次洗脱液重新加入吸附柱，室温放置1 min，12,000 rpm (13,680×g) 离心30 sec进行二次洗脱，或将 RNase-Free ddH₂O 置于65℃预热。
Introduction：
This kit utilizes an optimized buffer system suitable for total RNA extraction from various plant tissues, especially those rich in polysaccharides and polyphenols. It has a wide range of applications, and the extraction process does not require additional toxic or hazardous reagents such as phenol, chloroform, or β‑mercaptoethanol.
This kit is safe, efficient, rapid, and convenient. The RNA purified with this kit is of high purity, with minimal contamination from proteins, genomic DNA, and other impurities. Using this kit, dozens of micrograms of high‑purity RNA can be purified from 100–150 mg of plant material. The extracted RNA can be directly used in various molecular biology experiments, including Northern blot, dot blot, mRNA purification, in vitro translation, RNase protection assay, RT‑PCR, real‑time RT‑PCR, and cDNA library construction.
Precautions
Operators should wear disposable masks and gloves, change gloves frequently during the experiment, and use RNase‑free plasticware and tips to avoid RNase contamination.
This kit removes most genomic DNA contamination, allowing the RNA to be used directly in downstream experiments without DNase I digestion. However, nucleic acid content varies significantly among different samples or different input amounts. If downstream experiments require very high RNA purity, optional DNase I digestion can be performed (see step 5). DNase I can be purchased separately.
Fresh plant samples are recommended for RNA extraction. If extraction cannot be performed immediately, samples should be frozen in liquid nitrogen and stored below ‑70°C. Avoid repeated freeze‑thaw cycles, as this will affect RNA yield and quality.
Protocol
Preparation before use: Add 48 mL of absolute ethanol to Buffer RW2.
1. Homogenization
Add 600 μL Buffer RCL to a 2 mL centrifuge tube. Quickly grind plant leaves or fruit flesh into powder in liquid nitrogen. Weigh 50–100 mg of sample and transfer to the centrifuge tube containing Buffer RCL. Close the lid and shake vigorously about 20 times to mix the sample with RCL. Add 10 μL Proteinase K, immediately vortex for 30 seconds, and centrifuge at 14,000 rpm (18,630 × g) for 2 minutes.
Note:
If the sample has low RNA content, add 800 μL Buffer RCL and increase the sample amount to 100–130 mg.
For dry seed samples, add RE Reagent (available free of charge) to Buffer RCL before use.
2. Transfer ≤ 700 μL of the supernatant (the volume varies depending on the sample) to a Genomic DNA Removal Column DE (placed in a collection tube). Centrifuge at 14,000 rpm (18,630 × g) for 30 seconds. Discard the removal column and keep the flow‑through in the collection tube.
3. Measure the volume of the flow‑through (the volume may differ between samples; for the same tissue type from the same species, measuring one sample is sufficient). Add 0.7 volumes of Buffer RBE relative to the flow‑through volume (precipitation or flocculent material may appear – this is normal). Mix well, then transfer the mixture to a Spin Column RA (placed in a collection tube). Load no more than 700 μL per spin (if the mixture volume exceeds 700 μL, load in two separate spins). Centrifuge at 14,000 rpm (18,630 × g) for 30 seconds. Discard the flow‑through and return the column to the collection tube.
4. If not performing DNase I digestion, add 700 μL Buffer RW to the column. Centrifuge at 12,000 rpm (13,680 × g) for 30 seconds. Discard the flow‑through and return the column to the collection tube. Then proceed to steps 6–9.
5. DNase I digestion (optional)
If downstream experiments require very high RNA purity, optional DNase I digestion can be performed. If DNase I digestion is chosen, skip step 4 and directly follow steps 5–9.
5.1 Add 350 μL Buffer RW to the column. Centrifuge at 12,000 rpm (13,680 × g) for 30 seconds. Discard the flow‑through and return the column to the collection tube.
5.2 Preparation of DNase I stock solution: Dissolve the DNase I powder (2000 U) in 550 μL RNase‑Free ddH₂O. Mix gently, aliquot, and store at ‑30 to ‑15°C (can be stored for 9 months).
Note: Thawed DNase I stock solution can be stored at 2‑8°C for up to 6 weeks. Do not refreeze.
5.3 Preparation of DNase I working solution: Transfer 10 μL of DNase I stock solution to a new RNase‑Free centrifuge tube, add 70 μL DNase I Buffer, and mix gently.
5.4 Add 80 μL of the DNase I working solution to the center of the column. Let stand at room temperature for 15 minutes.
5.5 Add 350 μL Buffer RW to the column. Centrifuge at 12,000 rpm (13,680 × g) for 30 seconds. Discard the flow‑through and return the column to the collection tube.
6. Add 500 μL Buffer RCW to the column. Centrifuge at 12,000 rpm (13,680 × g) for 30 seconds. Discard the flow‑through and return the column to the collection tube.
7. Add 700 μL Buffer RW2 (ensure absolute ethanol has been added before use) to the column. Centrifuge at 12,000 rpm (13,680 × g) for 30 seconds. Discard the flow‑through and return the column to the collection tube.
8. Centrifuge at 12,000 rpm (13,680 × g) for 2 minutes.
9. Transfer the column to a new 1.5 mL RNase‑Free centrifuge tube. Open the lid and let stand at room temperature for 1 minute to completely air‑dry any residual ethanol. Add 30–100 μL RNase‑Free ddH₂O to the center of the membrane without touching it. Let stand at room temperature for 1 minute, then centrifuge at 12,000 rpm (13,680 × g) for 30 seconds. The obtained RNA can be used directly in downstream experiments or stored below ‑80°C.
Note: The volume of RNase‑Free ddH₂O should not be less than 30 μL; smaller volumes will reduce recovery. To increase RNA yield, reload the first eluate onto the column, let stand at room temperature for 1 minute, and centrifuge again at 12,000 rpm (13,680 × g) for 30 seconds (second elution), or preheat the RNase‑Free ddH₂O to 65°C.
规格
别名
多糖多酚植物总RNA提取试剂盒（柱式法）
规格或纯度
BioReagent,分子生物学级
英文名称
Universal Plant Total RNA Rapid Extraction Kit
应用
RNA Extraction
样品类型
植物
储存与运输
储存条件
室温
运输条件
常规运输
稳定性与储存
Store at room temperature long term (15 months).
组分与储存
| U1521342 | Component | 50T | Storage |
| U1521342A | Buffer RCL | 33 mL | RT. |
| U1521342B | Buffer RBE | 20 mL | RT. |
| U1521342C | Buffer RW | 38 mL | RT. |
| U1521342D | Buffer RCW | 28 mL | RT. |
| U1521342E | Buffer RW2 | 12 mL | RT. |
| U1521342F | RNase-Free ddH₂O | 15 mL | RT. |
| U1521342G | Proteinase K | 0.7 mL | RT. |
| U1521342H | 基因组清除柱DE (含2mL收集管) | 1 EA×50 | RT. |
| U1521342I | 吸附柱RA (含2mL收集管) | 1 EA×50 | RT. |
| U1521342J | RNase-Free离心管 (1.5mL) | 1 EA×50 | RT. |
本试剂盒所有组分可置于室温干燥条件下保存15个月。
Buffer RCL在储存时可能会形成沉淀，如果有沉淀出现，请于37℃加热溶解，然后待恢复至室温后使用。
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F0636935 |
分析证书 | U1521342 |
技术文档和文章
溶液计算器
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