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The EXTRACTME GENOMIC DNA kit is designed for the rapid and efficient purification of high quality genomic, mitochondrial, bacterial, parasite or viral DNA from solid tissues (fresh, frozen, formalin-preserved or paraffin-embedded), physiological fluids (urine, cerebrospinal fluid, peritoneal fluid, pleural fluid, sputum), fresh and frozen blood (human and mammalian), human and animal mucosa membrane swabs (including buccal, nasal, pharyngeal and vaginal swabs), semen, hair, rodent tails, insects, bacteria, yeast and cell cultures. The isolation protocol and buffer formulations have been optimized for high isolation efficiency and DNA purity. The product is intended for research use only.
- fresh or frozen solid tissue: 1-30 mg
- formalin-preserved tissue: 1-30 mg
- paraffin-embedded tissue: 1-30 mg
- physiological fluids (urine, PMR, peritoneal fluid): up to 5 ml
- cell culture: 103-107 cells
- broth or plate bacterial or yeast culture, frozen cell pellet
- buccal, nasal, pharyngeal, vaginal, blood and saliva swabs or semen
- fresh or frozen blood: up to 1 ml
- hair: 1-30 mg
- insects: 1-30 mg
The typical efficiencies of DNA isolation from fresh biological material are given in section XIII.
approx. 50 μg DNA
- approx. 12 minutes (lysis time not included)
- 10-60 minutes for sample preparation
A260/A280 ratio = 1.7 – 1.9
The DNA purification procedure consists of four steps and utilizes spin minicolumns with membranes which efficiently and selectively bind nucleic acids. In the first step, biological material is lysed by Proteinase K in optimized GL Buffer. At this stage all the cellular membranes and proteins are degraded. When it is necessary to remove RNA usage of the RNase A is recommended. After the addition of chaotropic salts, the lysate is applied to the purification minicolumn membrane and the DNA is bound. The two-step washing stage effectively removes impurities and enzyme inhibitors. The purified DNA is eluted using either a low ionic strength buffer (Elution Buffer) or water (pH 7.0-9.0) and may be used directly in all downstream applications such as PCR, quantitive real-time PCR, pharmacogenic research, Southern blotting, single-nucleotide polymorphism (SNP), short tandem repeat (STR) genotyping, DNA sequencing, enzymatic restriction, ligation and so forth, or stored until ready to use.
Quality control
The quality of each production batch (LOT) of EXTRACTME GENOMIC DNA kit is tested using standard QC procedures. The purified DNA concentration, quality and stability are evaluated by gel electrophoresis and spectrophotometer. In addition, the functional quality is tested by qPCR and digestion with restriction enzymes.