Transfection grade linear polyethyleneimine (MW40000)

Product Introduction

Transfection grade linear polyethyleneimine（MW40000）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, widely used in recombinant proteinsAAVWaiting for production.

andMW25000Compared to transfection reagents,MW40000The transfection reagent has good solubility and can be directly dissolved in water without the need to adjust the acidity or alkalinity of the solubilizing agent; The operation is simpler and the transfection effect is better.

Basic Information

Instructions for Use

Reagent configuration

Required materials:

Deionized water or above biological grade water1 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 polyethyleneimine（MW40000）Dissolve in900 mLStir deionized water evenly on a magnetic stirrer until dissolved;

⑵ use1 mol/L*AdjustmentpHto6.9~7.1(such aspHgreater than7.1Use hydrochloric acid topHAdjust the value to6.9~7.1）；

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 in4Can be stored at ℃1A month.

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 reagent complex:

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

⑵ Dilute an appropriate amount with suitable serum-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 reagent complexGently mix wellTransfection6-8After hours, replace with fresh preheated culture medium；

⑶ inAppropriate conditionsmiddleAfter 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.