SLS’s offer an optimized DNA extraction kit for plasmids using a modified alkaline lysis method. Unlike the extraction of genomic DNA, where all DNA is extracted, the isolation of plasmid DNA involves the separation of circular plasmid DNA from bacterial genomic DNA. Furthermore, the majority of the isolated plasmid DNA should remain supercoiled for increased efficiency in downstream applications such as transfection.
The scale of the plasmid DNA extraction kit used depends on the amounts of starting material and the yield of plasmid DNA that is required. Most kits come in mini, midi, and maxi prep sizes. The majority of applications use the mini kit size, mainly for screening. The midi kit is used for generating sufficient plasmid DNA for multiple manipulations or high-volume applications such as Southern blotting or pulsed field gel electrophoresis.
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Benefits
Minimize DNA loss with a kit that removes contaminants without precipitation or other handling steps that degrade DNA.
Improve reliability and reproducibility in downstream applications with a kit that removes RNA and other impurities that cause plasmid DNA to behave unpredictably.
Eliminate the use of hazardous organic compounds such as cesium chloride, phenol, chloroform, and ethidium bromide.
Format
Spin columns
Throughput
1–24 samples
Preparation time
24 minipreps in 30 minutes
Equipment required
Microcentrifuge or vacuum manifold
Lysate clearing
Centrifugation
Capacity of column reservoir
700 µl
Minimum elution buffer volume
50 µl
Culture volume for high-copy plasmids
1–5 ml
Culture volume for low-copy plasmids/cosmids
1–10 ml
Yield is variable and depends both on the particular E. coli strain used and the cell density of the bacterial culture.
Kit Contents
Storage Temperature
(upon receipt)
Spin Columns with collection tube
15-25°C
Buffer P1
15-25°C
Buffer P2
15-25°C
Buffer PN3
15-25°C
Buffer PW1
15-25°C
Buffer PW2§ (Concentrate)
15-25°C
Buffer PE
15-25°C
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers.
For Research Use Only. Not for use in diagnostic procedures.
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