EasyPure Blood Genomic DNA isolation Kit
EasyPure® Blood Genomic DNA Kit
EasyPure® Blood Genomic DNA Kit provides a simple and convenient way to isolate high quality genomic DNA from 5-250 μl of fresh or frozen blood. Whole blood is incubated with binding buffer to release DNA. DNA is bound to silica-based column. The isolated DNA is suitable for PCR, restriction enzyme digestion and southern blotting.
| Cat. No. |
EE121 | ||
| Speicification | With Rnase | EE121-01 | 50 rxns |
| EE121-02 | 200 rxns | ||
| No RNase | EE121-11 | 50 rxns | |
| EE121-12 | 200 rxns | ||
| Storage: |
RNase A and Proteinase K at -20°C for one year. Other components at room temperature (15-25°C) for one year |
||
| Shipping | RNase A on dry ice (-70 °C); others at room temperature. | ||
| Description | Simple and fast, red cell lysis buffer is no longer needed. | ||
| Complete removal of contaminants and inhibitors. | |||
| DNA yield up to 15 μg. • DNA range of 20-50 kb. | |||
| Column based purification, no organic extraction or ethanol precipitation. | |||
| Suitable for EDTA, sodium citrate and heparin-anticoagulated fresh and frozen blood in a volume of 5 μl to 250 μl. | |||
Yield of Extraction
For blood with nonnucleated erythrocytes, 5 μl~250 μl blood is recommended.
If not, 5 μl~20 μl is used.
|
Material |
Amount |
Genomic DNA |
|
Total Blood |
5 μl-250 μl |
≤ 10 μg |
Kit Contents
|
Component |
EE121-01 (50 rxns) |
EE121-02 (200 rxns) |
|
Binding Buffer 3 (BB3) |
30 ml |
110 ml |
|
Clean Buffer 3 (CB3) |
6 ml |
24 ml |
|
Wash Buffer 3 (WB3) |
12 ml |
2×22 ml |
|
Elution Buffer (EB) |
25 ml |
80 ml |
|
RNase A(20mg/ml) |
500 µl |
2×1 ml |
|
Proteinase K(20mg/ml) |
1 ml |
4×1 ml |
|
Spin Column with collection Tubes |
50 each |
2×100 each |
Procedures
Before starting, add the indicated volume of 96%-100% ethanol into the concentrated CB3 and WB3.
|
Component |
EE121-01 (50 rxns) |
EE121-02 (200 rxns) |
|
Clean Buffer 3 (CB3) |
24 ml |
96 ml |
|
Wash Buffer 3 (WB3) |
48 ml |
2×88 ml |
All centrifugation steps are carried out at room temperature.
1. Add the appropriate volume of blood, 20 μl Proteinase K and 500 μl BB3 into a microcentrifuge tube. Mix for 15 seconds by vortexing, and then incubate at room temperature for 10 minutes.
Optional: If RNA-free genomic DNA is required, add 20 μl RNase A before incubating.
2. Centrifuge briefly, and add all the lysate to a spin column. centrifuge at 12,000×g for 1 minute, discard the flow through.
3. Add 500 μl CB3 (check to ensure you have added ethanol), centrifuge the tube at 12,000×g for 30 seconds, discard flow through.
4. Add 500 μl WB3 (check to ensure you have added ethanol), centrifuge the tube at 12,000×g for 30 seconds, discard flow through.
5. Repeat step 4 once.
6. Place the spin column to a collection tube. Centrifuge the empty column at 12,000 rpm for 2 minutes to remove any residual WB3. Air-dry the spin column at room temperature for several minutes.
7. Place the spin column in a sterile 1.5 ml microcentrifuge tube. Add 50-200 μl of Elution Buffer (preheated to 60°C, able to increase DNA yield) or distilled water (pH >7.0) to the center of column. Incubate at room temperature for 1 minute. Centrifuge at 12,000 rpm for 1 minute to elute the genomic DNA (to recover more DNA, add EB or distilled water again to perform a second elution). For long-term storage, store the purified DNA at -20°C
Notes
• Do not use too many starting materials in case it affects the extraction performance
• To ensure the quality of extracted DNA, use fresh material and avoid repeated thawing and freezing. DNA quality depends on the types of material and storage time.
• Use sterile tubes and pipette tips to avoid the contamination from DNase
• You may perform the second elution step using the same microcentrifuge tube or different tubes
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