To help life science research,
let more research results benefit mankind
To help life science research,
let more research results benefit mankind
Endotoxin-free plasmids can be extracted from 5~15 mL of bacterial cultures, which is suitable for cell transfection and other routine plasmid experiments
Highlights
High purity: Use unique endotoxin precipitation technology to specifically remove endotoxins.
High-efficiency transfection: suitable for transfection of most cells, including endotoxin-sensitive cells.
Easy to operate: Using adsorption column technology to specifically adsorb plasmids, no filtration is required, and the operation is easier.
Wide application: can be used in animal and plant cell transfection and molecular biology experiments
Introduction
This kit uses an improved alkali lysis treatment method and silica membrane adsorption technology to obtain endotoxin-free high-purity plasmid DNA specifically and efficiently; the unique endotoxin precipitation technology requires no filtration and is easy to operate. This kit is suitable for extracting plasmids from 5 to 15 mL of overnight bacterial cultures. The resulting plasmids can be directly used in molecular biology experiments such as cell transfection, enzyme digestion, ligation, transformation, PCR, and sequencing
Composition
Contents | Amount |
RNase A (10 mg/mL) | 300 μL |
Buffer BL | 30 mL |
Buffer P1 | 30 mL |
Buffer P2 | 30 mL |
Buffer P4 | 30 mL |
Endotoxin Removal Buffer | 9 mL |
Buffer W1 | 24 mL |
Elution Buffer | 15 mL |
Adsorption Column EC (with Collection Tubes) | 50 |
Collection Tubes | 50 |
Related products
Type | Composition | Cat. No. | Amount | Introduction |
Nucleic Acid Electrophoresis | AG801 | 100 g | High purity and clear background | |
One Step Rapid Cloning | SC612 | 50 rxns | Different from the traditional seamless cloning kit, the kit can easily achieve 2~6 fragments seamless cloning. |
Q1:Why is plasmid DNA yield low?
A1:
1) Plasmid copy number is low, plasmid >10 kb or gram-positive bacterial plasmid. The amount of bacteria used should be increased, 300~500 mL of overnight culture can be used, and the eluent Buffer EB should be preheated in a 60°C water bath, and the adsorption and elution time can be appropriately extended to increase the extraction efficiency;
2) The problem of bacteria species. There is a loss of plasmids during the storage process of the strains. Before culturing the bacteria, it is recommended to streak and activate to stabilize the yield;
3) The bacteria are not fully lysed. Bacteria must be fully resuspended in Buffer I/RNase A or the bacterial cells should not be too large to avoid clumping or excessive bacteria that cannot be lysed and reduce the yield.
Q2:Why is there genomic DNA contamination in plasmid DNA?
A2:
1) The bacterial culture time is too long. The culture time of bacterial liquid should be controlled within 12-16 h;
2) The cracking problem. When adding Buffer II, it must be mixed by gentle inversion; when processing multiple samples, the total time should not exceed 5 min from the time of adding Buffer II.
Manual: Click to download
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