Column DNA Lego Kit, 200 izolations

Product description

Column DNA Lego Kit is a universal modular system (Lego-like) for pure DNA isolation from various sources. The method is based on capacity of silica surfaces to bind DNA in the presence of chaotropic agents (Proc. Natl. Acad. Sci USA, 76: 615-619, 1979).

When compared to other methods, DNA isolation with Column DNA Lego Kit is:

• 1. faster - does not require ultracentrifugation, ethanol precipitation and/or drying,

• 2. safer - does not require phenol extraction and/or use of ethidium bromide,

• 3. purer - no contamination with proteins, RNA, and/or their cleavage products,

• 4. simpler and cheaper - than isolation of DNA with popular DNA Lego Kit, which we introduced to the market more than a decade ago. DNA isolated using Column DNA Lego Kit can be used for variety of molecular-biology techniques such as sequencing, cloning, PCR, restriction enzymes cutting, mutagenesis in vitro, transcription in vitro, labeling reactions, transformation, or transfection.

Advantage of the Column DNA Lego Kit is that its components can be bought separately and universally used for isolation of (1) plasmid DNA, (2) DNA fragments from agarose gels, (3) DNA from solutions, including amplified DNA after PCR, and (4) genomic DNA.

Fig. 1. Comparison of plasmid DNAs isolated by three different methods. For plasmid DNA isolation from transformed E. coli cells alkaline lysis followed by neutralization was used. Then the plasmids were isolated by CsCl gradient ultracentrifugation (1, Ultrac. CsCl), commercial kit of a major producer of plasmid isolation kits (2, Producer X), or Column DNA Lego kit (3, Lego kit). Plasmids obtained were analyzed by electrophoresis in agarose gel without any prior treatment (-), or after partial cutting with HaeIII restriction enzyme (+). DNA in the gel was stained with ethidium bromide. The migration of DNA markers of 200-1500 bps (M) and untreated plasmids (PL) is indicated.

Conclusions: The results show that plasmids isolated with Column DNA Lego kit are comparable in quality to plasmids isolated by classical method of ultracentrifugation in CsCl gradient and that their yield is almost twice as much when compared to other column based kits of commercial producers. Higher yield is apparently related to the use of guanidine thiocyanate in Column DNA Lego kit, while other producers use cheaper guanidine chloride.      

Technical data

Unit definition

• Column DNA Lego Kit allows 200 isolations of pure DNA samples.

Storage

• At room temperature (+15 to +25^oC), except RNase which should be stored at -15 to -25^oC freezer and Suspension buffer [P1] with RNase stored at fridge (+2 to +8^oC) where it will be stable for 6 months. Storage of the Kit at fridge may lead to formation of salt precipitates in the buffers, which should disappear at room temperature. The Kit is shipped at ambient temperature.

Components

Column DNA Lego Kit (200 purifications):

• DNA bind columns (200 pcs) and collection tubes (200 pcs) in 4 boxes
• DNA bind buffer [L1], 60 ml
• Wash buffer [L2], 250 ml
• Elution buffer [L3], 20 ml
• Instructions 

Column DNA Lego Kit + plasmid supplement (200 purifications), possesses all components of the Column DNA Lego Kit and furthermore 4 solutions required for plasmid DNA isolation

• Suspension buffer [P1], 50 ml
• RNase A, DNase-free (10 mg/ml), 0.5 ml
• Lysis buffer - blue [P2], 50 ml
• Neutralization buffer [P3], 10 ml