SweTransRNA Transfection Reagent 0.5ml

Product Description/Introduction

This product is a high-performance cationic polymer-based RNA transfection reagent that delivers RNA directly to the cytoplasm of target cells without being degraded by nucleases. The gene expression process takes place in the cytoplasm without entering the nucleus and is not restricted by the transcriptional regulation of the target cell. This product has the advantages of little influence by serum, extremely low toxicity, good stability, easy operation and good repeatability. It can be applied to RNA transfection of various adherent cells and suspension cells.

Storage and Shipping Conditions

Ship with wet ice; Store at 2-8℃ for 12 months.

Product Content

Assay Protocol / Procedures

1. Pre-transfection preparation of adherent cells (6-well plate for example, other specifications refer to the attached table):

a. One day before transfection, the cells are planted uniformly in the well plate at a suitable density so that the cells converge to 70-85% during formal transfection;

b. Before transfection, remove the original cell culture medium, wash the cells with room temperature PBS buffer for 1-2 times, and then replace with 1.9 mL of serum-free or low-serum (less than 5%) basal medium (different cells have different serum dependency, please adjust the serum percentage as appropriate);

c. Cells are placed in a cell culture incubator after fluid exchange and transfection complexes are prepared;

2. Preparation of suspension cells for transfection:

a. On the day of transfection, after centrifuging the appropriate amount of cells to remove the medium, resuspend the cells in serum-free or low-serum (5% or less) basal medium to the appropriate concentration, and inoculate 1.9 mL of the resuspension uniformly in the well plate;

b. Place the well plate inoculated with suspended cells in a cell culture incubator and start to prepare the transfection complex (the volume ratio between the transfection complex and the culture medium can be adjusted appropriately by referring to the ratio of adherent cells);

3. Transfection complex preparation:

a. Prepare Solution A: 50 μL of serum-free basal medium with 80 pmol of RNA, blown up and mixed;

b. Preparation of solution B: 50 μL of serum-free basal medium with 4-20 μL of SweTransRNA transfection reagent, blown up and mixed;

c. Leave Liquid A and Liquid B at room temperature for 5 minutes;

d. Add solution B to solution A, gently blow to mix well, and continue to incubate at room temperature for 15-20 min to form the RNA-transfection reagent complex;

4. Transfection of cells:

a. The RNA-transfection reagent complex obtained in the previous step is added to the orifice plate with the liquid replaced in "Step 1" or "step 2";

b. After gently shaking the culture plate to induce the medium containing the transfection complex to mix well with the medium in the well plate, the well plate is incubated at 37°C in a 5% CO2 incubator;

c. After 4-6 h of incubation, the different cells are changed into the corresponding fresh medium and placed in the incubator for further incubation;

d. After 24-48 h of transfection, cells are observed or collected.

Note

1. Please use healthy cells in good condition, and resuscitated cells are recommended to be passaged at least twice before transfection.

2. Please use high quality RNA for transfection.

3. The product is used for the first time, it is recommended to pack separately to avoid repeated freezing and thawing.

4. Under the premise of cell tolerance, the incubation time of RNA-transfection reagent complex can be extended appropriately, and the transfection effect is better. The ratio of RNA to transfection reagents can be adjusted as appropriate.

5. For your health and safety, please wear lab coat and gloves during operation.

Table: The amount of transfection reagents and RNA required for different culture plates