Spin & Vacuum type of product for separating/ purifying high yield of Plasmid DNA (Midi Scale)
- Possible to separate and purify plasmid DNA effectively (Midi Scale).
- High copy number : 30~50 ml culture volume.
- Low copy number : 100 ml culture volume.
- Possible to separate and purify high yield and purified plasmid DNA within 30minutes.
- Possible to remove Cell lysate easily using Pre-filtration column.
- Ethanol precipitation is not necessary by using “Swing bucket rotor only” method.
- Separation/purification of plasmid DNA over 30 kb.
DNA-midi™ SV Plasmid DNA Purification Kit is used for purifying and separating high yield of plasmid DNA from midi scale of bacterial cell culture volume using silica membrane and optimized alkali lysis buffer system. It is composed of two column system including binding column(pre-filtration column and DNA binding membrane) in order to remove cell lysate easily and to perform separation/purification using vacuum method. (Figure 1.reference. Pre & Binding Column Combination)Neutralization buffering system used for pH neutralization and binding can perform not only separation/purification for plasmid DNA over 30Kb, but also remove chromosomal DNA, cell debris effectively after lysis. Also, pH change through alkaline lysis method can be observed visually in real time because the product contains lysis & neutralization indicator such as Lysis Viewer. Product is used for either High-copy plasmid DNA or low-copy plasmid DNA and in order to perform separation and purification effectively from endA+ strain(HB101, the JM strains, NM series strains, PR series strains, etc.), Washing Buffer A is provided separately.
Figure 1.Pre & Binding Column combination
|M1 Buffer(Resuspension Buffer)||60ml × 1 ea|
|M2 Buffer(Lysis Buffer)||60ml × 1 ea|
|M3 Buffer(Neutralization Buffer)||60ml × 1 ea|
|Washing Buffer A||150ml × 1 ea|
|Washing Buffer B||40ml × 1 ea
20ml × 1 ea
|Elution Buffer||30ml × 1 ea|
|Pre Column(Clear color)||25 Columns|
|Binding Column(Yellow color)||25 Columns|
|RNase A, lyophilized||40 mg x 1 tube|
Data Comparison for extraction efficiency based on size of plasmid DNA
After extracting various size of plasmid DNA using DNA-midiTM SV plasmid DNA Purification Kit, the results verified plasmid DNA over 40kb is extracted.
Lane M, 1kb ladder; lane 1, pUC18 (2.9 Kb); lane 2, pTA (7.1 Kb); lane 3, pCEP4(10.5 Kb); lane 4, pAdEASY-1 (33.2 Kb); lane 5, pJM17 (40.2 Kb)
Results for Transfection & Luciferase Assay
After performing separation/purification of pBl-GL plasmid DNA using DNA-midiTM SV plasmid DNA Purification Kit, transfected into 293T and K562 cell and conducted luciferase assay. The results verified that kit performed better compare to competitors.