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EXTRACTME DNA BACTERIA KIT

EXTRACTME DNA BACTERIA KIT
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The EXTRACTME DNA BACTERIA kit is designed for the rapid and efficient purification of high quality bacterial DNA from broth and plate cultures as well as frozen cells. The isolation protocol and buffer formulations were optimized for high isolation efficiency and DNA purity.
 
SAMPLE MATERIAL
broth or plate bacterial culture, frozen cell pellet
 
EFFICIENCY
- up to 5x 108 cells → 100%
- 1x 109 ÷ 3x 109 → 75-90% (when the modified protocol for isolation from large number of cells is followed)
- 1x 109 ÷ 3x 109 → ≤ 60% (when the standard isolation protocol is followed)
 
BINDING CAPACITY
approx. 40 μg DNA
 
TIME REQUIRED
40-60 minutes (including incubation time)
 
DNA PURITY
A260/A280 ratio = 1.7 – 1.9
 
Principle
The DNA purification procedure consists of four steps and utilises spin minicolumns with membranes which efficiently and selectively bind nucleic acids. In the first isolation step, the cell walls, membranes and proteins are degraded by lysis buffer and Proteinase K. To obtain an RNA-free DNA sample, the RNA is removed by RNase A. After the addition of chaotropic salts, the lysate is applied to the purification column membrane and the DNA is bound. The two-step washing stage effectively removes impurities and enzyme inhibitors. The purified DNA is eluted using 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, qPCR, Southern blotting, 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 DNA BACTERIA 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
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