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LNP常用细胞 RNA 转染试剂盒是一种基于脂质纳米粒(LNP)技术研发的新型即用型转染试剂,利用LNP 独特的跨膜转运和内体逃逸机制,实现对 RNA 高效递送,广泛适用于不同类型 RNA 在贴壁或悬浮的常用细胞中的转染。
易用:无需特定包封设备,一步混合即可实现 RNA-LNP 瞬时包封,无需孵育,可立即进行转染,转染后无需更换培养基。
高效:微量样品的包封率可达 80%以上,实现对常用细胞的高效转染。
安全:本品不含乙醇,采用新型生物可降解脂质材料,细胞毒性极低,且经 0.2μm 微孔滤膜过滤。
组分表
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运输与保存方法
受控常温(≤25℃)运输。2℃~8℃保存。不得冷冻!
适用范围
LNP常用细胞 RNA 转染试剂盒适用于绝大多数真核细胞 RNA 体外转染,其中 RNA 类型包括 siRNA、miRNA、gRNA、mRNA、saRNA 和 circRNA 等。
本产品仅供研究使用。
注意事项
(1) RNA 原液的最佳溶剂为无酶水,且 mRNA 浓度不低于 200ng/μL、短链 RNA 浓度不低于 100ng/μL、siRNA 浓度不低于 10μM,其他类型的溶剂对转染效果可能会有一定影响,具体情况可以联系我们。
(2) 制备 RNA 预混液请确保使用试剂盒专用 Buffer 稀释 RNA 原液,不得使用 PBS 或培养基。
(3) 请确保 RNA 预混液浓度为 40ng/μL。若多种 RNA 共转染,如 CRISPR Cas9 mRNA 与 gRNA 共转染,则 RNA 预混液浓度为 Cas9 mRNA + gRNA 的总浓度,Cas9 mRNA 与 gRNA 的推荐用量比为 1:1~1:2(μgmRNA:μggRNA)。
(4) RNA/LNP 的比例是 LNP 包封的关键因素,如需增减每孔用量,请按以下操作进行:
1、对于 mRNA 转染和 mRNA+gRNA 共转染,如需增减每孔 mRNA 用量,请按比例增加或减少加入细胞的每孔 RNALNP复合物的体积。
2、对于 siRNA 转染,如需增加每孔 siRNA 用量,请按比例增加加入细胞的每孔 RNA-LNP复合物的体积。
如需减少每孔 siRNA 用量,LNP转染试剂的最低用量应不低于推荐用量 II。以 96 孔为例,当 siRNA 每孔用量降为 0.5pmol 时,仅需调整 siRNA 预混液浓度为 0.5μM,转染试剂最低用量为 0.5μL/孔,其他操作维持不变。
(5) RNA-LNP复合物在室温条件下可稳定存放 8 小时,请勿将 RNA-LNP复合物置于冷冻条件。
(6) 转染前后均无需更换细胞培养基,含有血清、双抗的完全培养基对转染效果无明显影响;转染 6h 以后可按细胞常规培养流程,进行换液或其他操作。
附录 A:细胞转染操作说明
1. LNP常用细胞 RNA 转染试剂盒操作流程(推荐用量Ⅰ)

2. 不同培养容器中每孔试剂用量(推荐用量Ⅰ)
RNA 细胞转染步骤 | 试剂 | 96 孔板 | 24 孔板 | 6 孔板 |
1) 40ng/μL RNA 预混液制备 | Buffer(以 RNA 原液浓度 1μg/μL 为例) | 2.4μL | 12μL | 60μL |
| RNA | 0.1μg | 0.5μg | 2.5μg |
2) RNA-LNP复合物制备 | LNP常用细胞 RNA 转染试剂 | 2.5μL | 12.5μL | 62.5μL |
| RNA 预混液 | 2.5μL | 12.5μL | 62.5μL |
3) 加入细胞 | RNA-LNP复合物 | 5μL | 25μL | 125μL |
siRNA 细胞转染步骤 | 试剂 | 96 孔板 | 24 孔板 | 6 孔板 |
1) 1pmol/μL siRNA 预混液制备 (即 1μM) | Buffer(以 siRNA 原液浓度 10μM 为例) | 0.9μL | 4.5μL | 22.5μL |
| siRNA | 1pmol | 5pmol | 25pmol |
2) siRNA-LNP复合物制备 | LNP常用细胞 RNA 转染试剂 | 1μL | 5μL | 25μL |
| siRNA 预混液 | 1μL | 5μL | 25μL |
3) 加入细胞 | siRNA-LNP复合物 | 2μL | 10μL | 50μL |
3. LNP常用细胞 RNA 转染试剂盒操作流程(推荐用量Ⅱ)
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4. 不同培养容器中每孔试剂用量(推荐用量Ⅱ)
RNA 细胞转染步骤 | 试剂 | 96 孔板 | 24 孔板 | 6 孔板 |
1) 40ng/μL RNA 预混液制备 | Buffer(以 RNA 原液浓度 1μg/μL 为例) | 2.4μL | 12μL | 60μL |
| RNA | 0.1μg | 0.5μg | 2.5μg |
2) LNP预混液制备 | Buffer | 1.25μL | 6.25μL | 31.25μL |
| LNP常用细胞 RNA 转染试剂 | 1.25μL | 6.25μL | 31.25μL |
3) RNA-LNP复合物制备 | LNP预混液 | 2.5μL | 12.5μL | 62.5μL |
| RNA 预混液 | 2.5μL | 12.5μL | 62.5μL |
4)加入细胞 | RNA-LNP复合物 | 5μL | 25μL | 125μL |
siRNA 细胞转染步骤 | 试剂 | 96 孔板 | 24 孔板 | 6 孔板 |
1) 1pmol/μL siRNA 预混液制备 (即 1μM) | Buffer(以 siRNA 原液浓度 10μM 为例) | 0.9μL | 4.5μL | 22.5μL |
| siRNA | 1pmol | 5pmol | 25pmol |
2) LNP预混液制备 | Buffer | 0.5μL | 2.5μL | 12.5μL |
| LNP常用细胞 RNA 转染试剂 | 0.5μL* | 2.5μL* | 12.5μL* |
3) siRNA-LNP复合物制备 | LNP预混液 | 1μL | 5μL | 25μL |
| siRNA 预混液 | 1μL | 5μL | 25μL |
4) 加入细胞 | siRNA-LNP复合物 | 2μL | 10μL | 50μL |
*每孔 LNP常用细胞 RNA 转染试剂推荐用量为 siRNA 转染的最低用量。
附录 B:CRISPR/Cas9 mRNA+gRNA 共转染推荐方案
1. Cas9 mRNA+gRNA 共转染操作流程
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2. 不同培养容器中每孔试剂用量
Cas9 mRNA+gRNA 细胞转染步骤 | 试剂 | 96 孔板 | 24 孔板 | 6 孔板 |
1) 40ng/μL RNA 预混液制备 | Buffer | 4.8μL | 24μL | 120μL |
| Cas9 mRNA(以 mRNA 原液浓度 1μg/μL 为例) | 0.1μg | 0.5μg | 2.5μg |
| gRNA(以 gRNA 原液浓度 1μg/μL 为例) | 0.1μg | 0.5μg | 2.5μg |
2) LNP预混液制备 | Buffer | 2.5μL | 12.5μL | 62.5μL |
| LNP常用细胞 RNA 转染试剂 | 2.5μL | 12.5μL | 62.5μL |
3) RNA-LNP复合物制备 | LNP预混液 | 5μL | 25μL | 125μL |
| RNA 预混液 | 5μL | 25μL | 125μL |
4) 加入细胞 | RNA-LNP复合物 | 10μL | 50μL | 250μL |
LNP RNA Transfection Reagent (Common Cells) is a novel ready-to-use transfection reagent developed based on lipid nanoparticle (LNP) technology. Leveraging the unique transmembrane transport and endosomal escape mechanisms of LNPs, it enables efficient delivery of RNA and is widely applicable for transfecting various types of RNA into adherent or suspension conventional cell lines.
User-Friendly: No specialized encapsulation equipment required. RNA-LNP transient encapsulation is completed via one-step mixing with no incubation period; transfection can be performed immediately after preparation, and medium replacement is unnecessary post-transfection.
High Efficiency: The encapsulation efficiency of trace samples can exceed 80%, supporting robust transfection performance across common cell lines.
Low Cytotoxicity & Safety: This product is ethanol-free and formulated with novel biodegradable lipid materials, resulting in extremely low cytotoxicity. In addition, the reagent is sterile-filtered through a 0.2 μm microporous membrane.
Components List
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Transportation and Storage
Ship under controlled ambient temperature (≤25°C). Store at 2°C ~ 8°C. Do NOT freeze!
Scope of Application
This LNP RNA Transfection Reagent (Common Cells) is suitable for in vitro RNA transfection of the vast majority of eukaryotic cells. Applicable RNA types include siRNA, miRNA, gRNA, mRNA, saRNA, circRNA, etc.This product is for research use only.
Precautions
(1) Nuclease-free water is the optimal solvent for stock RNA solutions. The concentration of mRNA shall be no less than 200 ng/μL; short-chain RNA no less than 100 ng/μL; siRNA no less than 10 μM. Other solvents may compromise transfection efficiency. Please contact us for relevant details.
(2) Prepare RNA premix by diluting stock RNA with the dedicated Buffer supplied in this kit. Do not use PBS or cell culture medium.
(3) Adjust the final concentration of RNA premix to 40 ng/μL. For co-transfection of multiple RNAs (e.g. CRISPR Cas9 mRNA and gRNA), the concentration refers to the total concentration of Cas9 mRNA plus gRNA. The recommended mass ratio of Cas9 mRNA to gRNA is 1:1 ~ 1:2 (μg mRNA : μg gRNA).
(4) The RNA/LNP ratio is a critical parameter for LNP encapsulation. If adjusting the dosage per well, follow the instructions below:
1. For mRNA transfection and mRNA+gRNA co-transfection: if increasing or decreasing the mRNA dosage per well, proportionally adjust the volume of RNA-LNP complex added to each well.
2. For siRNA transfection: if increasing the siRNA dosage per well, proportionally increase the volume of RNA-LNP complex added to each well.
If reducing the siRNA dosage per well, the minimum volume of LNP transfection reagent shall not be lower than Recommended Dosage II. Take a 96-well plate as an example: when siRNA dosage is reduced to 0.5 pmol per well, adjust the siRNA premix concentration to 0.5 μM with a minimum reagent volume of 0.5 μL per well; all other steps remain unchanged.
(5) RNA-LNP complexes remain stable at room temperature for up to 8 hours. Do not freeze RNA-LNP complexes.
(6) Medium replacement is not required before or after transfection. Complete medium containing serum and antibiotics shows no significant interference with transfection results. Medium exchange or routine cell maintenance can be performed 6 hours post-transfection.
Appendix A: Cell Transfection Protocol
1. Operating Procedure for LNP RNA Transfection Reagent (Common Cells) (Recommended Dosage I)
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2. Reagent Dosage Per Well for Different Culture Vessels (Recommended Dosage I)
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3. Operating Protocol for LNP RNA Transfection Reagent (Common Cells) (Recommended Dosage II)
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4. Reagent Dosage Per Well for Different Culture Vessels (Recommended Dosage II)
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Appendix B: Recommended Co-transfection Protocol for CRISPR/Cas9 mRNA + gRNA
1. Operating Procedure for Co-transfection of Cas9 mRNA and gRNA
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2. Reagent Dosage Per Well for Different Culture Vessels
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | L1375584 |
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