BioReagent 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
避光,室温。常规运输 。请查阅批次 COA 获取详细规格。
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。
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。
RNA 的种类来源比较多，提取制备的方法各异，一般有苯酚法、去污剂法和盐酸胍法，其中最常用的是苯酚法，即 Trizol 法提取 RNA，异硫氰酸胍作为强的阴离子表面活性剂，可以有效的解离核蛋白与核酸的复合体，并对 RNase 产生强烈的抑制作用，保持 RNA 的完整性，加入氯仿等试剂并离心，上层清液用异丙醇沉淀回收总 RNA。
本品适用于从各种组织或细胞中快速分离总 RNA，既可用于小量样品(50～100 mg 组织、5×10⁶ 细胞)，也可用于大量样品(>1g 组织、>10⁷ 细胞)，提取的总 RNA 质量高，可用于 Northern Blot、Dot Blot、polyA筛选、体外翻译、RNase 保护分析和分子克隆，该试剂盒具有以下特点：①适用范围广；②操作简单，整个过程 90min 内完成；③纯度高；④污染少。
该试剂盒仅用于科研领域，不适用于临床诊断或其他用途。
组分表
|
自备材料
1、液氮、研钵或匀浆器、低温高速离心机、低温冰箱
2、经 RNase free 处理的移液器吸头、EP 管等耗材
3、无菌 PBS 缓冲液
操作步骤 (仅供参考)
1、实验准备：RNA 分离液室温放置 15min 确保两相分开，无菌 PBS 缓冲液、RNA 提取液和 RNA 沉淀液冰上预冷。
2、样品准备
1) 贴壁细胞：①直接裂解：直接在培养瓶 / 皿中加入 RNA 提取液裂解细胞，每 10cm² 面积加 1ml，用移液器吹打混匀。②胰蛋白酶消化：用无菌 PBS 洗涤细胞后，加入含有 0.05～0.25% 胰蛋白酶的 PBS 处理细胞，当细胞脱离容器壁后，加入含有血清的培养基终止反应，将细胞溶液转移至无 RNase 的离心管中，以下参考悬浮细胞相关操作步骤。
2) 悬浮细胞：收集 (1~5)×10⁶～10⁷动物、植物和酵母细胞或 10⁷细菌细胞加至 1.5ml 离心管，5000～6000g 离心 5min，用无菌 PBS 重悬细胞沉淀，再次 5000～6000g 离心 5min 收集细胞沉淀，去除上清，收集细胞时一定要将溶液去除干净，否则裂解不完全，降低 RNA 收获率，向沉淀中加入 0.5ml RNA 提取液，充分振荡混匀。
3) 组织：取新鲜动物或者植物组织或者 - 70℃冻存组织，50～100mg 组织在液氮中充分研磨或者加入 0.5ml RNA 提取液研磨或者用匀浆器匀浆处理，样品体积一般不超过提取液体积的 10%，研磨要迅速，以 1min 为佳。
4) 血液：取 0.5～1 ml 新鲜或冻存的血液，12000g 离心 5 min，去除血浆，加入 0.5ml RNA 提取液，充分振荡混匀。
3、核酸分离：将细胞液或匀浆液转移至 1.5ml 离心管，加入 1/10 体积的乙酸钠缓冲液，颠倒混匀 5~6 次，加入等体积 RNA 分离液，颠倒混匀 5~6 次后震荡 10‑20s，冰上放置 15min，使核蛋白与核酸完全分离。
4、样品分层：12000 g 4℃离心 10～15 min，RNA 在上层水相。
5、沉淀 RNA：吸取上层水相 (约 0.5ml) 转移至新的 1.5ml 离心管中 (不要吸取任何中间层物质，否则会有染色体 DNA 污染)，加入等体积 RNA 沉淀液混匀，冰上放置 10～15 min，12000 g 4℃离心 10 min，离心后管侧或管底形成胶状沉淀，弃上清。
6、洗涤 RNA：加入 1ml RNA 洗涤液轻轻洗涤沉淀，冰上放置 5～10 min，7500g 4℃离心 5 min，弃上清，室温干燥 10～30 min，不宜过分干燥，否则 RNA 难以溶解。
7、溶解 RNA：加入 30～50ul RNase‑free ddH₂O 充分溶解，‑70℃长期保存或直接用于后续试验，对于肝、胰腺、肾等组织中 RNase 含量高的样品沉淀用 RNA 保存液溶解。
注意事项
1、样品保存：加入 RNA 提取液混匀后样品可在‑70℃放置 1 月；RNA 样品可以在 70% 酒精中‑70℃保存 2～4 周；如果需要长期保存，应置于超低温冰箱中保存。
2、RNA 提取液和 RNA 分离液含有腐蚀性物质，污染皮肤或眼睛后，立即用清水或生理盐水冲洗，必要时寻求医生的帮助。
3、为了您的安全和健康，请穿实验服并戴一次性手套操作。
4、试剂开封后请尽快使用，以防影响后续实验效果。
有效期
12 个月有效。
There are diverse types and sources of RNA, and various extraction and preparation methods are available, including phenol‑based method, detergent method and guanidine hydrochloride method. Among them, the phenol‑based method, namely the Trizol method, is the most commonly‑used for RNA extraction. Guanidine isothiocyanate, a potent anionic surfactant, can effectively dissociate nucleoprotein‑nucleic acid complexes and exert strong inhibitory effects on RNase to preserve RNA integrity. After adding reagents such as chloroform followed by centrifugation, total RNA is recovered by precipitating the aqueous supernatant with isopropanol.
This product is designed for rapid isolation of total RNA from various tissues or cells. It is compatible with both small‑scale samples (50‑100 mg tissue, 5×10⁶ cells) and large‑scale samples (>1 g tissue, >10⁷ cells). The extracted total RNA is of high quality and can be applied in Northern Blot, Dot Blot, polyA selection, in‑vitro translation, RNase protection assay and molecular cloning. This kit features the following advantages: ① Wide application scope; ② Simple operation; the whole procedure can be completed within 90 min; ③ High purity; ④ Low contamination.
This kit is for research use only. It is not intended for clinical diagnosis or other applications.
Component
|
Reagents, consumables and Equipments not provided
1. Liquid nitrogen, mortar or homogenizer, refrigerated high‑speed centrifuge, low‑temperature refrigerator
2. RNase‑free pipette tips, EP tubes and other consumables
3. Sterile PBS buffer
Operating Procedures (For Reference Only)
1. Experimental preparation: Allow RNA separation solution to stand at room temperature for 15 min to ensure complete phase separation. Pre‑chill sterile PBS buffer, RNA extraction solution and RNA precipitation solution on ice.
2. Sample preparation
① Adherent cells:
Direct lysis: Add RNA extraction solution directly into culture flasks/plates to lyse cells at 1 mL per 10 cm² growth area, and mix well by pipetting.
Trypsin digestion: Wash cells with sterile PBS, then treat cells with PBS containing 0.05‑0.25% trypsin. Once cells detach from the vessel surface, quench the reaction by adding serum‑supplemented culture medium. Transfer the cell suspension into RNase‑free centrifuge tubes, and follow the procedures for suspension cells below.
②Suspension cells: Transfer (1‑5)×10⁶‑10⁷ animal, plant or yeast cells, or 10⁷ bacterial cells into a 1.5 mL centrifuge tube. Centrifuge at 5000‑6000 g for 5 min. Resuspend the cell pellet with sterile PBS and centrifuge again at 5000‑6000 g for 5 min to collect the pellet. Remove the supernatant completely; residual liquid will cause incomplete lysis and reduce RNA yield. Add 0.5 mL RNA extraction solution to the pellet and vortex thoroughly.
③Tissues: Use fresh animal/plant tissues or tissues stored at ‑70 ℃. Fully grind 50‑100 mg tissue in liquid nitrogen, or grind/homogenize with 0.5 mL RNA extraction solution. The sample volume should generally not exceed 10% of the extraction solution volume. Perform grinding rapidly, ideally within 1 min.
④Blood: Take 0.5‑1 mL fresh or frozen blood. Centrifuge at 12000 g for 5 min and discard plasma. Add 0.5 mL RNA extraction solution and vortex thoroughly.
3. Nucleic acid separation: Transfer cell lysate or tissue homogenate to a 1.5 mL centrifuge tube. Add 1/10 volume of sodium acetate buffer and mix by inverting the tube 5‑6 times. Add an equal volume of RNA separation solution, invert 5‑6 times, then vortex for 10‑20 s. Incubate on ice for 15 min for complete dissociation of nucleoprotein‑nucleic acid complexes.
4. Phase separation: Centrifuge at 12000 g, 4 ℃ for 10‑15 min. RNA resides in the upper aqueous phase.
5. RNA precipitation: Carefully transfer the upper aqueous phase (approx. 0.5 mL) to a new 1.5 mL centrifuge tube. Avoid aspirating any interphase material to prevent chromosomal DNA contamination. Add an equal volume of RNA precipitation solution and mix. Incubate on ice for 10‑15 min, then centrifuge at 12000 g, 4 ℃ for 10 min. A gelatinous pellet will form on the tube wall or bottom; discard the supernatant.
6. RNA washing: Add 1 mL RNA washing solution to gently wash the pellet. Incubate on ice for 5‑10 min, then centrifuge at 7500 g, 4 ℃ for 5 min and discard supernatant. Air‑dry at room temperature for 10‑30 min. Do not over‑dry, otherwise RNA will be difficult to dissolve.
7. RNA dissolution: Add 30‑50 μL RNase‑free ddH₂O to fully dissolve RNA. Store long‑term at ‑70 ℃ or proceed directly to downstream experiments. For RNase‑rich samples such as liver, pancreas and kidney tissues, dissolve the pellet with RNA storage solution.
Notes
1. Sample storage: Samples mixed with RNA extraction solution can be stored at ‑70 ℃ for 1 month. RNA samples can be stored in 70% ethanol at ‑70 ℃ for 2‑4 weeks. For long‑term preservation, store in an ultra‑low temperature refrigerator.
2. RNA extraction solution and RNA separation solution contain corrosive substances. In case of skin or eye contact, rinse immediately with plenty of tap water or normal saline and seek medical attention if necessary.
3. Wear laboratory coat and disposable gloves for personal safety and health during operation.
4. Use reagents promptly after opening to avoid compromised experimental performance.
Shelf‑life
Valid for 12 months
危险品化学品经营许可证（带存储）