Description

The EndoFree Maxi Plasmid Kit is based on silica membrane technology to extract endotoxin-free plasmid DNA. This kit provides a unique endotoxin removal process to generate high-quality plasmid DNA for downstream applications. It is designed to isolate plasmid DNA of the highest purity from 100-200 ml of bacterial culture and could yield up to 1.5 mg transfection grade plasmid DNA.

Key Features

• Fast and high yield: 200 μg-1.5 mg plasmid DNA yield in 1 hour with high proportion of super-coiled structure.

• High-purity: High-purity plasmid DNA are acquired by unique buffer system and Spin Column CP6.

• Excellent transfection efficiency: Suitable for transfection experiments of most cell lines.

• Wide range of applications: Suitable for restriction enzyme digestion, transformation, sequencing, microinjection, gene silencing and transfection experiments.

 

Specifications

Plasmid Type	Bacterial Culture Volume	Yield	Plasmids
Low Copy	200 ml	200-600 μg	pBR322, pACYC, pSC101, SuperCos, pWE15
High Copy	100 ml	500-1500 μg	pTZ, pUC, pBS, pGM-T

 

Components

1. RNase A (10 mg/ml): 500 μl
2. Buffer BL: 30 ml
3. Buffer P1: 100 ml
4. Buffer P2: 100 ml
5. Buffer P4: 100 ml
7. Buffer PW: 70 ml
8. Buffer TB: 30 ml
9. Filtration CS1: 10
10. Spin Columns CP6: 10
11. Collection Tubes 50 ml: 20

 

Storage

Store at room temperature.

 

Case Study

Low-copy plasmid pBR322, extracted from 200 ml bacterial culture using EndoFree Maxi Plasmid Kit, was elutedin 1 ml Buffer TB, with a concentration of 0.6 μg/μl.
High-copy plasmid pBS, extracted from 100 ml bacterialculture using EndoFree Maxi Plasmid Kit ,was eluted in 1ml Buffer TB, with a concentration of 1.2 μg/μl.

 

pEGFP obtained with EndoFree Maxi Plasmid Kit were separately transfected into endotoxin-insensitive cell line 293T and endotoxin-sensitive cell line Huh7 cells. Expression of GFP were detected in 24 hours after transfection.