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PEI 40K Transfection Reagent for cell biology Transfection Reagent

Specification:10ML
  • G1802-10ml
  • Servicebio
  • Servicebio

Product Description

G1802-10ml

Introduction:
PEI 40k Transfection Reagent (linearized polyethyleneimine transfection reagent) is a high-charge cationic polymer having a linear polyethyleneimine as a main body with a molecular weight of 40000, which belts positively charged, and can be effectively combined with the negative nucleic acid, forms a complex, and is introduced into the cell. Linearized polyethylene imine (PEI 40K) transfection reagent is low toxic, high transfection efficiency and low cost, is a very popular cell instantaneous transfection reagent. The currently proven linear PEI 40K transfection reagent is widely used in a variety of cell lines including HEK-293, HEK293T, CHO-K1, COS-1, COS-7, NIH / 3T3, SF9, HepG2 and HeLA cells, etc., transfection efficiency up to 80% ~ 90%.

 

Storage and transportation

Ice bag (Wet ICE) transportation; 4 ° C preservation, valid for 6 months.

Steps

1. Preparations before transfection of adherent cells (6-well plate as an example, refer to the attached table for other specifications):

a) The day before transfection, plant the cells evenly in the well plate at an appropriate density, and it is advisable for the cells to converge to 70-85% during the formal transfection;

b) Before transfection, remove the original cell culture medium, wash 1-2 times with PBS, and replace 1.8 mL serum-free or low-serum (less than 5%) basic medium, or Opti-MEM; different cells are serum-dependent Sex is different, adjusted according to specific cell types.

c) Place the replaced cells in the incubator and start to prepare the transfection complex.

2. Preparations before transfection of suspension cells:

a) On the day of transfection, centrifuge the appropriate amount of cells to remove the medium, and resuspend them in serum-free or low-serum (less than 5%) basic medium;

b) Place the prepared suspension cells in the incubator and start to prepare the transfection complex. The volume ratio between the transfection complex and the culture medium can be adjusted with reference to the ratio of adherent cells.

3. Preparation of transfection complex:

a) Prepare solution A: 100 μL of serum-free basal medium and 2 μg of plasmid DNA, mix well by pipetting;

b) Prepare solution B: 100 μL of serum-free basal medium and 6-8 μL of PEI 40 K Transfection Reagent, mix well by blowing and aspirating;

c) Add solution B to solution A, mix gently by pipetting, and incubate at room temperature for 15 minutes to form a DNA-PEI transfection complex.

4. Transfected cells:

a) Add the transfection complex obtained in the previous step by drawing a cross or a circle to the well plate that was previously replaced;

b) Hold the well plate and draw an "8" on the flat surface or gently shake the culture plate in other ways to make it fully mixed, and incubate in a 37°C, 5% CO2 incubator;

c) The DNA-PEI transfection complex is incubated with the cells for 4-6 hours to have better transfection efficiency. Prolonged or overnight incubation will have better transfection efficiency.

Precautions

1. Please use healthy cells in good condition. It is recommended that the recovered cells be passaged at least 2 times before transfection.

2. Please use high-quality, endotoxin-free plasmids.

3. High serum levels during transfection and the use of antibiotics may affect transfection efficiency and cell status.

4. The tolerance and sensitivity of different cells are different. The length of the transfection incubation time and the ratio of PEI/DNA use can be adjusted according to the situation.

5. This product is only used for scientific research, not for humans.

6. Please wear lab coats and disposable gloves during operation.

 

Attached table: Transfection usage of different cell culture vessels (for reference only)

 

Culture vessel

Growth area

(Cm2)

Number of inoculated cells

DNA amount

(ug)

PEI

(μL)

Transfection complex volume(μL)

total capacity

(mL)

96well plate

0.3

(2-4)×104

0.1

0.3-0.4

10

0.1

24well plate

1.9

(1.2-2.4)×105

0.5-1

1.5-4

50

0.5

12well plate

3.8

(2.4-4.8)×105

1-2

3-8

100

1

6well plate

9.5

(6-10)×105

2-4

6-16

200

2

10 cmPetri dish

55

(4-6)×106

12-24

36-96

1000

12

T75Culture flask

75

(6-10)×106

18-36

54-144

1000

15

This product is for scientific research purposes only, not for clinical diagnosis!

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