SweTransDNA Transfection Reagent 0.5ml

Product Description/Introduction

This product is a new generation of DNA transfection reagent with excellent performance of cationic polymer type, which has superb DNA binding and delivery ability, and can be used for transfection of plasmid DNA. This product has the advantages of very low toxicity, good stability, simple and easy transfection, good reproducibility, etc. It can be used for transient transfection and stable transfection of many types of 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 60-85% 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 2 μg of plasmid DNA, blown up and mixed well;

b. Preparation of solution B: 50 μL of serum-free basal medium with 2-10 μL of SweTransDNA transfection reagent was blown and mixed;

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

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

4. Transfection of cells:

a. The DNA-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, endotoxin-free plasmid DNA.

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 DNA-transfection reagent complex can be extended appropriately, and the transfection effect is better. The ratio of DNA 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 DNA required for different culture plates