Transfection grade의 plasmid 혹은 cosmid DNA을 10 mg까지 분리 정제할 수 있습니다
CsCl 농도구배 원심분리 두 번 한 것과 같은 동일한 순도
Transfection grade plasmid DNA 의 재현성 있는 yield.
Ethidium bromide, phenol, chloroform, 또는 CsCl 불필요
경제적인 preparation이 가능함
QIAGEN Plasmid Kits provide gravity-flow, anion-exchange tips for purification of transfection-grade plasmid DNA. The QIAGEN Plasmid Giga Kit delivers DNA yields of up to 10 mg. Lysate clearing and isopropanol precipitation are achieved by centrifugation.
Buffers P1, P2, P3, QBT, QC, QF, RNase A; for 100 plasmid mini-, 25 midi-, or 10 maxipreps
Transfection efficiency versus plasmid purification method.
Different pRSVcat DNA preparations using the methods indicated were introduced into the indicated cell lines by liposome-mediated transfection, and the efficiencies determined by measuring CAT expression levels after 40 h. Each bar represents the mean of 4 independent transfections (2 transfections with each of 2 independent plasmid preparations). The highest transfection efficiency was achieved with QIAGEN Plasmid Kits.
QIAGEN Plasmid Kit procedures.
Neutralized bacterial lysates are cleared by centrifugation. The cleared lysate is then loaded onto the anion-exchange tip where plasmid DNA selectively binds under appropriate low-salt and pH conditions. RNA, proteins, metabolites, and other low-molecular-weight impurities are removed by a medium-salt wash, and ultrapure plasmid DNA is eluted in high-salt buffer. The DNA is concentrated and desalted by isopropanol precipitation and collected by centrifugation.
The QIAGEN Plasmid Giga Kit uses gravity-flow QIAGEN-tip 10000 anion-exchange tips for efficient purification of plasmid DNA. Up to 10 mg high-copy plasmid DNA is purified from 2.5–5 liters culture. (Culture volumes depend on plasmid copy number, size of insert, host strain, and culture medium.) Plasmid DNA purified with QIAGEN Plasmid Kits is highly suitable for use in applications such as transfection (see figure "Transfection efficiency versus plasmid purification method"), cloning, and in vitro transcription.
The unique anion-exchange resin in QIAGEN-tips is developed exclusively for the purification of nucleic acids. Its exceptional separation properties result in DNA purity equivalent or superior to that obtained by two successive rounds of CsCl gradient centrifugation. Prepacked QIAGEN-tips operate by gravity flow and never run dry, minimizing the hands-on time required for plasmid preparation. The entire QIAGEN plasmid purification system avoids the use of toxic substances such as phenol, chloroform, ethidium bromide, and CsCl, minimizing hazard both to the user and the environment.
With QIAGEN Plasmid Kits, bacterial lysates are cleared by centrifugation. The cleared lysate is then loaded onto the anion-exchange tip where plasmid DNA selectively binds under appropriate low-salt and pH conditions. RNA, proteins, metabolites, and other low-molecular-weight impurities are removed by a medium-salt wash, and pure plasmid DNA is eluted in high-salt buffer (see flowchart "QIAGEN Plasmid Kit procedures"). The DNA is concentrated and desalted by isopropanol precipitation and collected by centrifugation.
Plasmid DNA purified with QIAGEN Plasmid Kits is highly suitable for use in applications, such as:
In vitro transcription
Transfection, cloning, sequencing, capillary sequencing etc.