PRINCIPLE

DNA purification procedure utilizes spin microcolumns with membranes which efficiently and selectively bind nucleic acids. In the first step of the clean-up protocol CB Buffer is added to a DNA sample. It causes proteins to degrade and enables DNA binding to the column membrane while in the gel-out protocol the DNA fragments is excised from an agarose gel and incubated in GB Buffer, which enables gel fragment solubilization and protein degradation. As an added convenience, the binding buffers contain a colour indicator, which facilitates easy monitoring of the solution’s pH for optimal DNA binding. The two-step washing stage efficiently removes impurities and enzyme inhibitors. Purified DNA is eluted with the use either a low ionic strength buffer (Elution Buffer) or water (pH 7.0–9.0) and can be used directly in all downstream applications such as PCR, qPCR, Southern blotting, DNA sequencing, enzymatic restriction, ligation and so forth or stored until ready to use.

PRODUCT SPECIFICATIONS

SAMPLE MATERIAL

CLEAN-UP: DNA sample
GEL-OUT: agarose fragment of up to 150 mg  containing DNA

YIELD

Depending on DNA fragment length (in the range  of 100 bp – 10 kb): 70–90 %

DNA FRAGMENT LENGTH

100 bp – 10 kb
DNA fragments in the 50–100 bp and 10–30 kbp range can also be purified, as can genomic and plasmid DNA, however the efficiency will be  decreased.

BINDING CAPACITY

Approx. 6 μg DNA

TIME REQUIRED

approx. 10 min for clean-up procedure
approx. 15 min for gel-out procedure

DNA PURITY

A²⁶⁰/A²⁸⁰ ratio = 1.7 – 1.9