EasyTrans Reagent

 

Cat. No. QYR026a Size: 0.8 mL

 

Cat. No.QYR026b Size: 1.5 mL

 

Store at +4℃ or -20℃

 

(Avoid repeated freeze-thaw cycles)

 

Description

EasyTrans has been validated to effectively and reproducibly transfect single siRNA or co-transfect DNA/siRNA to variety of mammalian cells with maximal transfection efficiency and minimal cytotoxicity. In addtion, because of its high transfection efficiency, no or minimal toxicity and immunogenicity, EasyTrans also is a superior gene delivery tool into many animals in vivo including mice, rats, tadpoles and ducks via intravenous, intraventricular, subcutaneous, tracheal and intraperitoneal injections. 

 

in vitro transfection

 

Use the following procedure to transfect mammalian cells in a 6-well format. For other formats, please see table 1.

 

1.Adherent cells: Cells should be seeded a day prior to transfection in 6-well plates at an appropriate density. Before transfection, cells should be washed twice with PBS, then cultured in 2 mL of basal medium (containing no additives ie serum, antibiotics or other proteins).

Suspension cells: Just prior to preparing complexes, plate cells at an appropriate density in 2 mL of basal medium (containing no additives ie serum, antibiotics or other proteins).

 

2.Dilute 4 μg DNA in 250 μL PBS buffer and mix by pipetting up-down. 

 

3.Dilute 10 μL DNAHitrans in 250 μL PBS, mix by pipetting up-down gently and incubate for 5 min at room temperature. Then combine the diluted DNA with diluted DNAHitrans, mix immediately by pipetting up-down and incubate for 20 minutes at room temperature.

 

4.Add the 500 μL of mixture to each well dropwise. Mix gently by rocking the plate back and forth.

 

5.Incubate cells at 37℃ in a CO2 incubator for 4-6 hours and then the medium is replaced by complete medium.

 

in vivo Transfection

 

1.Dilute 10 μg of DNA in 50 μL of sterile PBS solution. Vortex gently and spin down briefly. 

 

2.Dilute 5 μL of DNAHitrans in 50 μL of sterile PBS solution. Vortex gently and incubate for 5 min at room temperature.

 

3.Add the diluted 50 μL of DNAHitrans to the diluted 50 μL of DNA. Mix immediately by pipetting up-down immediately and spin down briefly. Incubate 20 min at room temperature.

 

4.Inject animals. Please see table 2. 

 

 

Table 1. Scaling Up or Down Transfections

Culture vessel	Surface area per well (cm2)	Volume of plating medium	DNA in PBS volume	DNAHitrans in PBS volume
96-well	0.3	100 μL	0.2 μg in 25 μL	0.5 μL in 25 μL
24-well	2	500 μL	0.8 μg in 50 μL	2.0 μL in 50 μL
12-well	4	1 mL	1.6 μg in 100 μL	4.0 μL in 100 μL
35-well	10	2 mL	4.0 μg in 250 uL	10 μL in 250 μL
6-well	10	2 mL	4.0 μg in 250 uL	10 μL in 250 uL
60-mm	20	5 mL	8.0 μg in 0.5mL	20 μL in 0.5 mL
10-cm	30	15 mL	24 μg in 1.5 mL	60 μL in 1.5 mL

                         

                                                                             Table 2. Suggested Amount of DNA and Maximum Injection Volume 

Animal	Route of injection	Suggested amount of DNA (μg)	Maximum injection volume (μL)
Adult mouse	Intravenous	25-125	400-600
	Retroorbital	40	200
	Intraperitoneal	100	600
	Heart	50	200
	Lung instillation	20	300
	Subcutaneous tumor	10	100
Nude mouse	Intravenous	50	200
	Subcutaneous tumor	10	100