Q1: Does this in vivo transfection reagent have bone tissue targeting?
There is no targeting. The particles formed by transfection reagents and RNA are at the nanoscale. Our in vivo distribution experiments have found that the RNA levels are the highest in the liver and lungs. This one is now a universal in vivo transfection reagent. The targeted products mentioned earlier are not in stock and need to be customized. Because we need to make chemical modifications. The laboratory has completed the in vivo targeted transfection reagents for tumor tissue, cartilage tissue and bone tissue. Some customers need to customize them, and the target heads of the three types are different. Cartilage targeting is to target chondrocytes. Bone targeting is to target osteoblasts
Q2: Must the RNA used be purified by PAGE and HPLC? Are there other purification methods for siRNA and miRNA?
When customers generally need RNA transfection reagents, it is for the study of RNA functions. Usually, single pure RNA molecules are used, and the main sources are prepared by chemical synthesis, semi-chemical synthesis or biosynthesis methods. The purification methods commonly used are PAGE or HPLC. Total RNA is obtained from biological samples, and a specific RNA molecule (a single RNA) needs to be further isolated. The amount of RNA extracted biologically is small and the types are numerous. The subsequent separation and purification methods need to be very complex and are generally not suitable for RNA transfection experiments.
Q3: 所使用的RNA必须使用PAGE和HPLC纯化方式么？siRNA和miRNA还有其他纯化方式么？
客户一般需要用到RNA转染试剂的时候是对RNA功能进行研究，一般使用单一纯品RNA分子，主要来源就是化学合成、半化学合成或者生物合成法制备的，所用纯化方法通常就是PAGE或者HPLC。从生物样本中进行总RNA获得，具体某个RNA分子（单一RNA）是需要进一步分离的。生物提取RNA量小，种类多，后续分离纯化方法需要很复杂，一般不适合做RNA转染实验。
Q4: Are the purification methods for siRNA and miRNA different from mRNA purification?
The RNA purification methods are all similar and there is no difference
Q5: siRNA和miRNA的纯化方式和mRNA纯化方式是否不同？
RNA纯化方式都类似，没有不同
Q6: Can the RNA and transfection reagent complex be diluted with other reagents?
It is recommended to use the isotonic solution tested in the laboratory (5% glucose solution prepared with DEPC sterile water) to prepare the RNA and transfection reagent complex. If other solutions need to be tried, DEPC sterile water or serum-free medium can be tested. However, the results were more stable when 5% glucose solution (w/v, prepared with sterile DEPC water) was selected.
Q7: RNA和转染试剂复合物可以用其他试剂稀释吗？
优先推荐实验室测试过的等渗溶液（DEPC无菌水配置的5%葡萄糖溶液）配置RNA和转染试剂复合物，如需尝试其他溶液，可测试DEPC无菌水或无血清培养基。但选用5%葡萄糖溶液（w/v，用无菌DEPC水配制）结果稳定性更好。
Q8: Why is glucose solution needed for dilution? To reduce injection volume, can the glucose diluent be omitted?
The dilution buffer recommended here is mainly to maintain an isotonic environment, keep the osmotic balance inside and outside the cells, and help maintain normal cell function and physiological state. After the non-isotonic solution is added, the smaller the volume, the lower the risk of damage. However, the final effect can only be further analyzed and judged by the customer based on the actual situation
Q9: 请问为什么需要使用葡萄糖溶液进行稀释？为了减少注射体积，可以不用葡萄糖稀释液吗？
这里推荐的稀释缓冲液，主要是为了维持等渗环境，保持了细胞内外的渗透平衡，有助于维持正常的细胞功能和生理状态。非等渗溶液加入后，体积越少，损害风险越小，但最终效果只能客户根据实际情况进一步分析判
Q10: 两款RNA转染试剂都可以做RNA和DNA共转么？
我们的RNA转染试剂（KX0110049&KX0110050）只针对RNA(包括siRNA、miRNA、重组RNA，不包括mRNA)，通过实验发现不适合DNA转染。因此也不适合RNA和DNA共转染。此外，目前市面上的试剂也是DNA和RNA转染试剂分开居多，一般都不能通用。
Q11: What is the difference between the [RNA In Vitro Transfection Reagent] and the [RNA In Vivo Transfection Reagent]?
The structure of the key raw material polymer is consistent, but the production process of the "RNA in vivo transfection reagent" has been improved, with increased purity and lower endotoxin to meet the requirements of in vivo experiments.
Q12: 【RNA体外转染试剂】和【RNA体内转染试剂】有什么区别？
关键原料聚合物的结构一致，但【RNA体内转染试剂】生产工艺上做了改进，纯度上提升，内毒素更低，以满足体内实验的需求。
Q13: Is your transfection reagent GMP grade?
no
Q14: For the siRNA in vivo transfection reagent, the product Q&A mentions non-targeting, most obvious in lung and liver. The customer's target organ is the spleen. Do you have relevant data for this?
There is no data on the spleen for the time being
Q15: How to verify efficiency?
(Note: It is recommended that regardless of whether precipitation is found or not, the prepared complete culture medium should be fully inoculated and cultured for pre-experiment detection. Usually, the test cells and the complete culture medium should be placed in a 37 ° C incubator overnight for observation.)" "
Q16: What factors affect knockdown efficiency?
① Small RNA sequence design: Efficient knockdown depends on the specificity and stability of small Rnas. ② High-purity RNA is a key factor for successful transfection. The content and purity of RNA should be determined before transfection (OD260/280 > 1.8). ③ Delivery method and dosage: The amount of RNA used should be determined based on the animal model, administration route, and target organ. The recommended proportioning dosage for RNA transfection in rodents is provided in the instruction manual. ④ Target tissues: Some tissues (such as the liver) are more likely to achieve high-knockdown due to their rich blood flow, while others (such as the brain) may require additional optimization.
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