Transfection grade linear polyethyleneimine (MW25000)

Transfection grade linear polyethyleneimine（MW25000）It is a polyethylene imine based transient transfection reagent product with high transfection efficiency. Polyethyleneimine is an organic macromolecule polymer with high cationic charge, which easily binds to negatively charged nucleic acid molecules to form positively charged complexes. The complex binds to cell surface anions and enters the cell through endocytosis. This transfection reagent has low cytotoxicity and is suitable for various cell lines. It has been widely used in recombinant proteinsAAVWaiting for production.

1. Reagent configuration

Required materials:

Deionized water or above biological grade water12 mol/LHydrochloric acid（HCl）The10 mol/L*（NaOH）One time use0.1~0.2Mm PESVacuum sterile filter, sterileHDPEOr polypropylene storage bottles.

Storage liquid configuration:

⑴ Translate into English1 gTransfection grade linear polyethyleneimine25,000dissolved in900 mLDeionized water, while stirring (can be heated to)60-80Add dropwise at ℃12 MHydrochloric acid, adjust topH＜2.0, Stir3More than hours until dissolution;

⑵ use10 M*AdjustmentpHto6.9~7.1；

⑶ Transfer the solution into a measuring cylinder (or cup)Inside, add waterconstant volume to1 L；

⑷ Use disposable0.2 µmFilter vacuum filtration for sterilization, i.e. obtain1 mg/mLThe storage liquid;

⑸ Can be divided into storage solutions as needed and stored in-20Can be stored at ℃1Year; After thawing, the storage solution cannot be refrozen.

2. Transfection operation process

Inoculate cells: To improve transfection efficiency, it is recommended to inoculate cells one day before transfection to maintain the cell density at the time of transfectionin70~80%As appropriate；

prepareDNA-Transfection reagentcompound:

⑴ Translate into EnglishDNAThe transfection reagents and diluents should be restored to room temperature first;

⑵ Dilute an appropriate amount with a suitable protein free culture mediumDNA；

⑶ According to a certain proportion(Every1 μg DNAneed to use1-5 μLTransfection reagent, it is recommended to explore the ratio by yourself)Add the diluted transfection reagent dropwise to the containerDNATest tube for solution(Do not reverse the order of addition)On one sidegentlyVortex, thoroughly mixLet it stand at room temperature10~25minuteincubation.

Transfected cells:

⑴ When transfected under serum-free conditions, remove the cell culture medium and replace it with serum-free culture medium;

⑵ Add an appropriate amount to the cellsDNA-Transfection reagentcomplexGently mix wellTransfection6-8After hours, replace with fresh preheated culture medium；

⑶ in37℃The5% CO₂incubatormiddleAfter cultivation and transfection24hourThe expression of the transferred gene can be detected (the appropriate detection time can be determined by oneself).

Transportation and storage conditions

Transportation conditions: room temperature transportation

Storage conditions: Powder at room temperature or4℃ storage, expiration date2Year.

Precautions

1. For most cells, every1 μg DNAuse3 μLTransfection reagents can achieve high transfection efficiency, and it is also possible to try each method1 μg DNAuse1.5~4 μLOptimize the transfection reagent.

2. For your safety and health, please wear lab coats and disposable gloves when operating.

3. This product is only for scientific research purposes.