How do you make polyethyleneimine solution?

How do you make polyethyleneimine solution?

Preparation of PEI Stocks

1. Dissolve 100 mg in 100 mL sterile ddH2O.
2. Stir while slowly adding HCl to pH 7.0.
3. Mix for 10 minutes and then recheck pH.
4. Filter sterilize through 0.22 um filter.
5. Aliquot 500 uL to 1000 uL and store in -80C.
6. Thawed solution can be stored at 4C for up to 2 months, label a tube when thawed.

How do you dilute PEI?

Stir the PEI in MQ (1mg/ml) and dissolve by heating to ~80 degrees C. Vortex intermittently. It should go completely into solution in a relatively short amount of time.

How do you dissolve PEI Max?

Dissolve PEI in endotoxin-free dH2O that has been heated to approximately 80 °C . Let cool to room temperature. Adjust pH until it reaches pH 7 , filter sterilize (0.22 uM filter), aliquot, and store at -20 °C .

How do you make PEI Max?

In 1L glass beaker, suspend 1g of PEI MAX 40K in 900 mL water. 2. Add PTFE-coated stir bar to 1L glass beaker and set stirring to produce small vortex.

How do I make a 1mg ml PEI?

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1. Dissolve the powder in H,O. Adjust pH to 7.0 with HCl (the solution becomes clear as the pH is adjusted).
2. Filtrate through 0.22 um membrane.
3. Make 0.2~1ml aliquots, and store at —80°C.
4. For every batch of PEI, Test the transfection efficiency using different.
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What is PEI transfection?

DNA can be introduced into a host cell by transfection with polyethylenimine (PEI), a stable cationic polymer (Boussif et al., 1995). PEI condenses DNA into positively charged particles that bind to anionic cell surfaces. Our laboratory uses PEI over other cell transfection reagents because of its low cost.

What does PEI do in transfection?

DNA can be introduced into a host cell by transfection with polyethylenimine (PEI), a stable cationic polymer (Boussif et al., 1995). PEI condenses DNA into positively charged particles that bind to anionic cell surfaces.

What is transfection method?

Transfection is the process of introducing nucleic acids into eukaryotic cells by nonviral methods. Using various chemical or physical methods, this gene transfer technology enables the study of gene function and protein expression in a cellular environment.

Can PEI Max be frozen?

PEI-HCl-40K at 1mg/ml working solution: stable at 4 degree for up to 6 months. For long- term storage, put 3-5ml aliquots in -80 or -20 degree. Once thawed completely, filter through 0.22um filter, aliquot and keep at 4 degree. Do not freeze again.

Can PEI be frozen?

Background: Polyethylenimines (PEIs) are synthetic, charged polymers which function as transfection reagents based on their ability to compact DNA into complexes. Furthermore, in contrast to other polyethylenimines, PEI F25-LMW-based DNA or siRNA complexes allow freeze/thawing and frozen storage for several months.

What is PEI Max?

Polyethylenimine “Max” (PEI MAX) is a powerful, trusted, and cost-effective reagent widely considered as a current gold standard for both in vitro and in vivo transfection. PEI MAX has a high density of protonatable amino groups, with amino nitrogen as every third atom.

How do you resuspend someone on PEI?

METHOD

1. Pour ~450 mL of Milli-Q H2O into a 500-mL glass beaker.
2. Add 500 mg of PEI to the beaker with stirring.
3. Add concentrated HCl drop-wise to the solution to pH <2.0.
4. Stir until the PEI is dissolved (~2-3 h).
5. Add concentrated NaOH drop-wise to the solution to pH 7.0.
6. Pour the solution into a 500-mL glass cylinder.

What is polyethylenimine (PEI)?

Polyethylenimine (PEI) is a cationic polymer that contains repeating units composed of an amine group and a CH 2 CH 2 spacer. Notably, PEI is an organic polymer that has the highest positive charge density potential as every third atom of this polymer is a protonable amino nitrogen.

How do you prepare Pei stock solution?

PEI (linear MW 25,000) PEI stock solution (1 mg/mL) can be prepared as follows: 1. Dissolve the PEI powder in Milli-Q H 2 O. The solubility of PEI in water is low. Adjust the pH to 7.0 with 1 M HCl (the solution becomes clear as the pH is adjusted).

What is the molecular weight of poly(ethyleneimine) solution?

Concentrations of polymer solutions (such as PEI) are commonly reported either as a percentage or mg/mL, not in terms of molarity. The product description of Product P3143, Poly (ethyleneimine) solution, on sigmaaldrich.com appears to give two molecular weight values, Mn and Mw.

What is the percentage of Pei in the solution?

This product is nominally a 50% w/v solution of PEI in water, meaning 50 grams of poly (ethyleneimine) per 100 mL of solution. Concentrations of polymer solutions (such as PEI) are commonly reported either as a percentage or mg/mL, not in terms of molarity.

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