Search
Search
View additional product information for Invivofectamine™ 3.0 Reagent - FAQs (IVF3005, IVF3001)
19 product FAQs found
Isolating high-quality RNA is critical to the overall success of in vivo experiments.
Invivofectamine 3.0 Reagent is designed to be delivered slowly and evenly; a 200-µL injection should take between 10 to 20 seconds.
Sterile technique is vital to successful completion of in vivo experiments. All materials need to be sterile before injecting the complexed siRNA into animals.
The protocol is robust but specific. Protocol modifications will likely result in less than optimal performance.
Both positive and negative controls are recommended when performing any siRNA experiment. We typically use Invitrogen in vivo siRNA against Factor VII with Invivofectamine 3.0 Reagent, as a positive control for experiments. It is also good practice to use PBS solution as a negative control.
We recommend injecting 3-5 mice per treatment to assess the reproducibility of the results within an experiment.
To ensure the best results and minimal toxicity, it is very important to determine the weight of each animal to provide the correct dose and to maintain uniform dosing of multiple animals. The recommended dose for mice is 1 mg/kg ( IV) to start and reduce the dose depending on the potency of your siRNA.
We recommend the use of inbred strains for in vivo experiments, because these strains provide a uniform experimental background in which to determine gene knockdown and downstream effects. In addition, we recommend purchasing animals that are in good health from a qualified vendor. Starting with sick or lethargic mice is likely to result in problems during the experiment
Yes! We recommend that you test at least 3 different siRNA duplexes in vitro at several different concentrations and use the siRNA duplex with the highest potency for your in vivo experiments. When moving from in vitro to in vivo experiments, it is also important to double check the species specificity of the duplex.
Your siRNA must be at least 1.2 mg/mL in RNase/DNase-free water before complexing. siRNA stock concentrations lower than 1.2 mg/mL will result in low encapsulation efficiencies. We also recommend that you confirm the siRNA concentration before complexing with Invivofectamine 3.0 Reagent to verify the correct concentration. After resuspension in RNase/DNase-free water, measure the OD at 260 nm of several serial dilutions of the stock and calculate the concentration.
We recommend that your siRNA is of the highest quality available. In vivo purity siRNA from Invitrogen or highly purified and desalted siRNA is required for good results. Failures in synthesis, excess salts, free nucleotides, and improperly purified siRNA may result in poor performance in vivo.
No, "10 reactions" means it is for 10 injections into mice.
Find additional tips, troubleshooting help, and resources within our Transfection Basics Support Center.
Sorry, we have not tested this. We only have validated this reagent in-house for vivo delivery of siRNA and miRNA to the mouse liver by tail vein injection.
For mRNA in vivo delivery, please check out our Vivofectamine delivery solutions.
Find additional tips, troubleshooting help, and resources within our Transfection Support Center.
Keep the siRNA concentration at 2.4 mg/mL and follow protocol instructions for complex preparation, keeping the siRNA to Invivofectamine 3.0 Transfection Reagent ratio at 1.2:1 (µg:µL).
Find additional tips, troubleshooting help, and resources within our Transfection Support Center.
Invivofectamine 3.0 Reagent is an improved formulation with the following increased benefits:
- Lower toxicity
- Higher transfection efficiency with less siRNA; sustained knockdown up to 2 weeks
- Increased stability
- Shorter overall workflow; no dialysis or diafiltration needed
The Invivofectamine 3.0 Reagent-siRNA complexes may be used without the 6-fold dilution, in concentrated form. However, the complexes should be either dialyzed or diafiltrated before use.
Invivofectamine 3.0 Reagent-siRNA complexes may be stored up to two weeks at 4°C.
The Invivofectamine 3.0 Reagent has been validated in-house only for in vivo delivery of siRNA and miRNA to the mouse liver by tail vein injection.
You may perform up to 10 injections of 200 µL each of a 0.1 mg/mL siRNA complex solution (e.g., 20 g mouse, 1 mg (siRNA)/kg (body mass)).